Showing papers in "Acta Scientiae Veterinariae in 2012"

Relationship between Mineral Composition of Seminal Plasma and Semen Quality in Various Ram Breeds

Abstract: Background: Seminal plasma of mammals is a complex fluid, which serves as a carrier for the spermatozoa on their journey from the male testes to their target...

Antibacterial activity of essential oil of cinnamon and trans-cinnamaldehyde against Staphylococcus spp. isolated from clinical mastitis of cattle and goats

Abstract: Background: Milk production plays a role as a source of protein and constitutes an important socioeconomic factor for small farms. The lack of planning of the creators, together with the lack of specialized technical assistance, and the precarious hygienic and sanitary management, are barriers that hinder the development of the activity. Bovine and caprine mastitis is an infl ammation process of the mammary gland caused by microorganisms. Staphylococcus spp. is the main causative agent of mastitis, with higher prevalence in cases of clinical and subclinical manifestations. The most common treatment is based on intramammary infusion of antibacterial agents. However, cure rates obtained with such drugs are not always effectives, because it may determine the emergence of resistant bacteria as well increase amounts of antibiotic residues in milk. Nevertheless, the treatment of bovine subclinical mastitis caused by Staphylococcus spp. in the lactation can be economically unviable. Alternative treatments of bovine mastitis with plant derived compounds have been described. The investigation of its antimicrobial activity against bacterial agents of mastitis is justifi able. Materials, Methods & Results: This study evaluated the antimicrobial activity of essential oil (EO) of Cinnamomum zeylanicum (cinnamon) and the fraction trans-cinnamaldehyde (TC) against 65 isolates of Staphylococcus spp. from mastitis of cows (n = 32) and goats (n = 33). The cinnamon EO was extracted from the leaves and was composed majority by isoeugenol (96%), while the TC was acquired commercially. The antimicrobial susceptibility tests were performed according to CLSI M31-A3 agar diffusion method. Based on these tests, the isolates were divided into subgroups according to the resistance profi les. Subsequently, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by broth microdilution technique CLSI M31-A3. The MICs (geometric mean, GM) for cinnamon EO were 2032 mg/L and 2102 mg/L, respectively, for cattle and goats, while the majority fraction TC was most active with MICs (GM) of 598 mg/L and 875 mg/L, respectively, for cattle and goats. Discussion: Some differences between these results and those reported by other authors can be attributed to the composition of OEs or techniques employed. In this study, we used the broth microdilution technique based on the document M31-A3, from CLSI, for the determination of MICs and MBCs. The lack of an internationally standardized technique for evaluating OEs and plant extracts that allow different protocols undermines the comparisons of results. The results of this study confi rmed the antimicrobial activity of cinnamon EO and TC against clinical Staphylococcus spp. independent of the antibacterial susceptibility profi le. The best antimicrobial activity was observed with TC than in the cinnamon EO, which deserves further research investigating the use of these compound in the treatment of mastitis. The TC can be found in high concentration in cinnamon bark. However, the essential oil used in this work was extracted from the leaves, and the major component was isoeugenol. The cinnamon EO composition can be varied depending on the part of the plant where it is extracted.

Perdas econômicas das principais causas de condenações de carcaças de frangos de corte em Matadouros-Frigoríficos sob Inspeção Federal no Rio Grande do Sul, Brasil

Abstract: Background: A considerable fraction of the chicken carcass is condemned in slaughterhouses in Brazil, following the legal requirements by monitoring the Inspection Service of the Ministry of Agriculture, Livestock and Supply (MAPA) and Quality Control of agribusinesses. These losses result from partial or complete sentences, the fi rst being the most signifi cant. This study aimed to analyze the main causes of convictions and total post-mortem inspection of carcasses of broilers from slaughterhouses under SIF, as well as the analysis of the economic losses arising out of total convictions that occurred from January 2009 to June 2011. Materials, Methods & Results: This work was carried out by survey data generated by the Federal Inspection Service in a slaughterhouse located in southern Brazil, slaughtering on average 188.000 poultry.day-1, producing a 360 T.day-1 products. The data were collected from records of the occurrence of partial and total condemnation of carcasses of broilers slaughtered in the period January 2009 to June 2011, following the requirements approved by the Ethics Committee on Animal Use of the Federal University of Rio Grande do Sul (CEUA / UFRGS). For calculations of economic losses, it was considered the sum of poultry completely discarded and the production cost of chicken every six months of 2009, 2010 and 2011, with an average carcass yield of 2.5 kg chicken (cold carcass weight), a value of R$1.71. kg-1, R$1.44.kg-1 and R$1.77, respectively. Statistical analysis was performed using analysis of variance and comparisons between means using the Tukey test (5%) from the SAS Enterpise Guide (2003). During the period January 2009 to June 2011, the slaughter fridge 137.721.990 chickens slaughtered, of which 0.65% (893.869) were totally condemned and 4.74% (6.521.745) were partially condemned, and the main causes of contamination were found total convictions, cachexia and look disgusting. Making up a six-monthly review, it is observed that, regardless of the year under review, between the fi rst and second half there was no signifi cant difference (P > 0.05) in the percentage of total convictions for these diseases. Simulating the impacts caused by economic doom total, we obtained a R$ 585.830.94 loss, and R$ 755.294.40 R$ 842.522.72 in the first half of 2009, 2010 and 2011, in order. These values imply an annual growth of 28.93% between the fi rst year and 11.55% from 2010 to 2011, equivalent to a total economic loss of 43.81% over the past two years. Discussion: Ordinance No. 210, Annex IX, defi nes the destination and judging criteria of birds, will be subject to condemnation of carcasses post-mortem inspection with: abscesses, sacculitis, looks disgusting, cachexia, infection, injury, skin diseases, special, scalding excessive delayed evisceration, fractures, thin, infl ammation, sepsis syndrome and ascites tumors. Slaughtered poultry in the fi rst half of 2009, 0.54% (1.357.519) were totally condemned. In 2010, there was an increase in the percentage of animals condemned totally in the previous year, however, was not evidenced in the following year (2011) presented a stability in the percentage of birds in relation to convicted totally slaughtered birds. To minimize fractures and bruises at the time of harvesting and transport, integration with producers should be focused on awareness of Good Practice loading, transporting, unloading and animal welfare, which generate sentences in later stages of the process especially in the wings, thighs and breast.

Stress Response in Dairy Cows Related to Rectal Examination

Abstract: Background: Currently, rectal palpation is the cheapest and most convenient method which is using for detecting most of internal diseases, gynecologic conditions and artifi cial inseminations in dairy cows However, rectal palpation procedure is probably painful, resulting in physiological and behavioral stress reactions The life and metabolic activity can be affected by the oxidative stress which is one of the stress-related negative outcomes and characterized by the accumulation of radical oxygen species Many factors such as stress, diet, antibiotics and husbandry practices effect animals’ health and growth performance The aim of the study was emphasized to the effects of rectal palpation related to stress response and oxidant /antioxidant status in dairy cows Materials, Methods & Results: The study was conducted on ten 3-4 years old, non-pregnant Holstein dairy cows The rectal palpation procedure was performed in a closed area using the traditional method Blood samples were taken from each animal, an hour before and just after the rectal palpation procedure for detecting white blood cell, red blood cell, hemoglobin levels, hematocrit value, total antioxidant status, total oxidant status and nitric oxide, glucose and total cholesterol levels, as well as the plasma cortisol concentrations WBC, RBC, Hct and Hb values were measured from whole blood by using an automatic blood count device Plasma TAS and TOS levels were determined using a commercially available kit Nitric oxide decomposes rapidly in aerated solutions to form stable nitrite/nitrate products Plasma nitrite/ nitrate concentration was measured by a modifi ed method of Griess assay Plasma cortisol concentrations were determined using an ELISA kit Glucose and total cholesterol levels were analyzed in serum by using an auto-analyzer White blood cell levels and hematocrit values were signifi cantly increased just after the rectal palpation procedure compared to the baseline values (P < 005) whereas no signifi cant difference was found in red blood cell levels and hemoglobin concentrations Serum cholesterol concentrations have decreased after rectal palpation, serum glucose and cortisol concentrations have signifi cantly and markedly increased (P < 005), the relevant variation factors were 1341% and 2326% respectively Moreover, a remarkable and signifi cant increase in the circulating TOS compared to the baseline values (P < 005 variation factor 3178%) was observed after rectal palpation In parallel, the nitric oxide concentrations were also signifi cantly increased after rectal palpation procedure (P < 005 variation factor: 1351%) On the other hand, the total plasma antioxidant status was depressed and the variation intensity remained weak (variation factor: -724%) Discussion: In the present study the results indicate that the rectal palpation procedure will be considered as a stressful condition, as evidenced by the strong increases in circulating cortisol, TOS and NOx concentrations associated with decreases in TAS concentrations In addition, increased plasma cortisol concentrations in response to rectal palpation could be an additional factor responsible for the oxidative stress amplifi cation A more comprehensive identifi cation of the physiological changes during rectal palpation could be benefi cial for further researches in terms of accurate management practices in cattle practices and industry Moreover, the determination of the oxidative stress parameters could provide novel approaches for the evaluation the stress in cattle

Parasitismo do ácaro Ornithonyssus bursa em humanos no Sul do Brasil

Abstract: Background: The mite Ornithonyssus bursa is a bloodsucking parasite of domestic and wild birds and it is typical of tropical regions. On the host it can cause considerable loss of blood and lesions with intense itching. Cases of parasitism in human beings have been reported causing dermatitis, and most of these cases are due to the presence of nests of synanthropic birds near of the residences. When in large infestations or lack of hosts, the mites leave the nests to perform the parasitism in humans. Human infestation occurs mainly in people who handle infected birds and nests, or when happen the dispersion of these mites, through some path which lead to the interior of homes, factories and hospitals. Case: The present case occurred in November of 2008, when one of the dwellers of an apartment on the fourth floor of a building downtown in Santa Maria, woke up with pruritic lesions over the arms and legs. Upon inspection of the bedroom, several mites were seen, measuring almost a millimeter long, coming out of a pipe of a portable air conditioner installed on site. They were already scattered by the inside walls, floor and bed. The mites were collected and maintained in 70% alcohol for later identification. After the clarification the slides mounting were performed, and the mites with measure between 0.48 mm wide by 0.74 mm long were classified by the position of the anal opening, formed by the dorsal plate and three pairs of bristle on the sternal plate, as belonging to the species Ornithonyssus bursa. On external outlet of the air portable conditioner, a nest with a couple of birds and three cubs was found. The birds were identified as belonging to the species Myiodynastes maculatus, popularly known as brindle Bem-Te-Vi. The birds were removed from the place and treated with antiparasitic macrolactones to control the infestation. Discussion: In the present case report the O. bursa was found inside of an apartment in the downtown area, parasitizing birds which had their nest sheltered in the outlet air of a portable air conditioner. The presence of synanthropic bird brindle Bem-Te-Vi and the ease of access to the interior of the residence led to a home infestation of this ectoparasite. This fact may be happend due to the nesting of birds in urban buildings, seeking for a favorable environment for their survival, specially near of local that providing heat, like heaters, furnaces and hot water pipes. When the young birds parasitized by hematophagous leave the nests, the mites can cause the parasitism in humans mainly by food shortage. In the present study the occurrence of accidental home infestation and accidental parasitism in humans is related to the presence of many specimens of birds in the same nest, with offering of food and favorable environmental conditions for reproduction of the mites and their dissemination. Mite infestations in humans are characterized by erythematous papulovesicles distributed throughout the body, mainly on the limbs and chest. In this case report the parasitism caused erythematous lesions, especially in the arms with intense itching. The dermatitis produced by mesostigmatas hematophagous mites are not specific impeding the diagnosis, and may be confusing with other ectoparasites. For this reason the present diagnosis was performed by isolation and identification of the mite. Based on the description above it was possible to conclude that the bird Myiodynastes maculatus are parasitized by the mite Ornithonyssus bursa and its synanthropic habits allowed the infestation of mites in this residence.

Exposure to Leptospira spp. in Sick Dogs, Shelter Dogs and Dogs from an Endemic Area: Points to Consider*

Abstract: Background: Leptospirosis is a zoonosis caused by pathogenic spirochetes of the genus Leptospira. Rodents play an important role as maintenance hosts, but dogs can be signifi cant reservoirs for human infection in tropical areas as well as the source of disease outbreaks. Manifestations of disease in dogs vary from asymptomatic carriers to severe clinical signs and death. This study compared leptospiral exposure in dogs suspected to have leptospirosis and presented at a Veterinary Teaching Hospital (VTH), dogs from a Control Center of Zoonoses (CCZ) and dogs from a neighborhood with a high prevalence of human leptospirosis. Also, clinical signs, laboratory abnormalities and environmental risk factors associated with the infection were investigated at a population level and in a case-by-case approach. Materials, Methods & Results: Between May 2007 and February 2009, 253 dogs from Porto Alegre, Brazil, were enrolled in the study. Three populations were evaluated including dogs from and endemic area to human leptospirosis, dogs from a CCZ and dogs presented to a VTH. All dogs’ owners from the endemic area and from the VTH answered a questionnaire including dog’s information such as breed, age, vaccination status, environment, contact with other domestic animals, presence of rodents in the household, clinical signs, medications and if owners had leptospirosis diagnosed in the previous two years. The investigation of the exposure to pathogenic leptospires was based on serology using the Microscopic Agglutination Test (MAT), and polymerase chain reaction (PCR) using two sets of primers to detect pathogenic leptospiras in blood (leptospiremia) and urine (leptospiruria). Positive results were found in the three populations. The most prevalent serovars were Canicola, Icterohaemorrhagiae and Copenhageni, independent of the dog health condition. Leptospiruria occurred in 20.0%, 8.4% and 30.3% of CCZ, endemic area and VTH dogs, respectively. There was no association between seropositivity and leptospiremia or leptospiruria. The presence of rats in the environment was associated with leptospiruria (P = 0.02). Complete blood count (CBC), serum biochemistry (alanine aminotrasferase and creatinine) and urinalysis were also performed. Although increased serum creatinine (P = 0.009), jaundice (P = 0.004) and glucosuria (P = 0.04) were associated with leptospiruria in the VTH dogs, the absence of clinical signs or clinicopathologic alterations did not exclude the infection, as observed in several dogs from CCZ and from the endemic area. Discussion: As expected, the VTH showed the relatively highest percentage of positive samples (serology, leptospiruria and leptospiremia), since these were clinical cases. However, no statistical differences were found in the percentage of leptospiremia between VTH and the dogs from endemic area, neither in the percentage of leptospiruria or serology between VTH and dogs from CCZ. The most common serovars identifi ed by MAT were consistent with the fi ndings of other studies involving dogs in Southern Brazil. If creatinine is elevated, particularly if jaundice is present, the likelihood of leptospiral infection must be considered; however, normal fi ndings for these parameters do not rule out this diagnosis. Most of the dogs vaccinated the year before showed leptospiremia and/or leptospiruria, suggesting infection with a serovar not included in the vaccine, vaccine ineffi cacy or a wrong dog vaccination schedule. Subclinical infection is a problem when considering animals with leptospiruria that will likely remain untreated. The control of the environmental dissemination of pathogenic Leptospira spp. in urban settings should include the identifi cation of asymptomatic dogs.

Relationship between Levels of Peripheral Blood Testosterone, Sexual Behavior, Scrotal Circumference and Seminal parameters in Crossbred Rams

Abstract: Background: Measurements of testosterone, scrotal circumference and libido have a great value as indicators of onset of puberty, total semen production, semen quality, control of spermatogenesis, testicular state, pathological conditions of testes and the potential sub-fertility or infertility. Therefore, the aim of this study was to determine relationship between levels of peripheral blood testosterone, sexual behavior, scrotal circumference and seminal parameters in crossbred rams. Materials, Methods & Results: Twelve crossbred and sexually mature rams consisting¬ of three Arkharmerino × Moghani (AM × MG), three Baluchi × Moghani (BL × MG), and three Ghezel × Baluchi (GH × BL) and three Ghezel × Arkharmerino (GH × AM) were used in this study. The scrotal circumference (SC) was measured at monthly intervals. Every two weeks the 12 crossbred rams were evaluated for the degree of libido using three ovariectomized ewes. Sexual behavior of crossbred rams was evaluated in terms of (1) reaction time for the fi rst and second ejaculate (2) time taken for the fi rst and second ejaculate (3) number of mounts for the fi rst and second ejaculation. Ram semen was collected by artifi cial vagina and blood samples were obtained via jugular vein. Soon after the collection, semen characteristics and testosterone plasma were assayed. For estimation of the relationship between genetic group and other indices, Statistical analyses were performed by One-way ANOVA and Bivariate correlation coeffi cient was used to calculate the correlations between testosterone by spermatozoa parameters and libido in each genetic group; P < 0.05 considered signifi cant. The results showed that there were no signifi cant differences between crossbred rams in terms of semen characteristics except for spermatozoa progressive motility, semen volume and pH. The highest level of plasma testosterone was recorded in GH × BL rams (7.12 ± 1.87 ng/mL) and the lowest was for AM × MG genetic group (2.99 ± 1.90 ng/mL). However, any signifi cant difference wasn’t observed among the four genetic groups about plasma testosterone. At the present study the mean values of scrotal circumference of Arkharmerino × Moghani were higher than other genetic groups (P < 0.05). Also scrotal circumference showed a positive correlation with reaction time for the second ejaculation, semen pH and spermatozoa concentration (P < 0.05). Discussion: The sexual activity of males is infl uenced by test conditions and the methods applied in tests can vary considerably, even within the same experiment for example with regard to test duration and number of males and females. Among libido indices, only a signifi cant correlation was observed between reaction time for second ejaculate and scrotal circumference. A non-signifi cant correlation was observed between plasma testosterone, concentration, and semen traits. These differences between various reports could be due to the season of the study, breed, age of rams and many other environmental factors. It could be concluded that testosterone profi les is not a notable factor for estimating the quality of crossbred rams spermatogenesis and sexual activity. In addition to the above points, the males should be examined for a number of different tests have been used to assess the fertility or performance of crossbred rams, including scrotal measurement, semen examination, libido testing, hormonal profi le and the other examinations.

Avaliação do desempenho reprodutivo de vacas de corte lactantes submetidas à IATF a partir da aplicação do GnRH, da manifestação estral, da reutilização de dispositivos intravaginais e da condição corporal

Abstract: The success of fi xed-time artifi cial insemination (FTAI) is linked to several factors that affect pregnancy rates. The use of different hormones, estrus ex...

Somatic Cell Count and California Mastitis Test as a Diagnostic Tool for Subclinical Mastitis in Ewes

Abstract: Background: Infectious mastitis has been described as one of the main diseases affecting animals during lactation. The disease in sheep has been studied for many years in countries where mastitis has an economical importance. However, the interest in mastitis studies in animals raised for food production has increased, because the disease may cause a reduction in weight and an increased mortality in lambs. In this study, Somatic Cells Count (SCC) and California Mastitis Test (CMT) were related to bacterial isolation for mastitis diagnosis in Corriedale sheep. Material, Methods & Results: Twenty nine (29) ewes, varying from 2-5 years of age, with different numbers of lactation and born lambs and never before machine-milked were used. Milking was done from October to November, once a week in the morning period, with oxytocin application. Four milk collections were made, at biweekly intervals for bacteriologic, SCC and CMT test, which were analyzed considering each gland as a sampling unit. Low incidence of subclinical mastitis (9.66%) was observed, with the majority (15/17) caused by coagulase negative Staphylococcus. There was no change in milk production related to SCC or bacteria isolation. However changes in milk components occurred in the presence of subclinical mastitis. A poor relation (k = 0.115) was determined between the results obtained in the bacteriological test and the SCC, with low sensitivity (13.33%) and an increased number of false negative results (13%). Comparing the SCC and CMT results as the diagnostic method for subclinical mastitis, a low (r = 0.2319) but significant (P = 0.0209) correlation was observed, as well as a poor concordance (k = 0.152). Using bacterial isolation as the standard test, it was determined that the CMT has low sensitivity (28.57%) as the diagnostic method of mastitis in ewes. Discussion: Mastitis has been considered an economically important disease in the production of sheep for meat and wool. According to reports, the frequency of its clinical occurrence may range from zero to 50%. In meat-producing herds, a low weight gain in lambs has been associated with subclinical mastitis and the study of mastitis in Corriedale sheep is justified since this is considered a breed of meat sheep with the best milk production. This breed is being crossed with milkproducing breeds, such as Laucane, to form milk-producing herds. Bacterial isolation has been adopted as the diagnostic method of mastitis in all livestock breeds. Similarly to the observation made by this study, Staphylococcus and, in a few cases, Streptococcus, have been the microorganisms most frequently involved in subclinical mastitis in sheep. The milk from ewes with mastitis tends to have a lower fat and lactose content than that of healthy ewes, due to the affected secretory function of these animals. The SCC of milk ewes has not yet been established, but its count in a healthy udder may reach up to 1.5x10 6 cel.mL -1 . Similarly, the CMT score to be used in sheep is still controversial, but the maximum score (+++) is adopted to indicate mastitis. The high number of false-negative and false-positive reactions observed in SCC and CMT tests means that healthy and ill animals are incorrectly identified and that no preventive and curative measures are adopted. Since the utilization of only one diagnosis method in sheep mastitis, without confirmation by bacteriologic test is not conclusive, the SCC and CMT should be used cautiously in sheep mastitis diagnosis.

Morphology and involution of the yolk sac during early gestation bovine (Bos indicus)

Abstract: Background: The bovine yolk sac derives from visceral endoderm and its development occurs between days 18-23 of gestation. The study of this membrane is important for comparative data and has already been performed in rodents, sheep and in cattle, especially Bos taurus. In species Bos indicus the yolk sac has not quite been studied and is believed that there are morphological differences between these species. The yolk sac undergoes a process of involution and degeneration during embryonic development and none vestige of it is found in late gestation. The period in which occurs the involution of the yolk sac coincides with the period of increased pregnancy loss in cattle, and changes in the morphology of this membrane may indicate the reasons for such high loss rates. Thus, considering that the yolk sac is important for embryonic circulation and metabolic transmission, besides participating actively in the process of cattle placentation, this study aimed characterize morphologically the involution of the bovine yolk sac. Materials, Methods & Results: The early gestational period was determined between days 20 and 70 post-insemination (p.i), according to the exterior characteristics of embryo/fetus. For macroscopic analyzes the uterus was dissected to expose the fetal membranes and subsequently the embryo/fetus was photographed. The samples were fi xed for light microscopy and transmission electron microscopy. The yolk sac that emerges from the ventral part of the embryo was prominent and composed by a central part with two thin peripheral projections of different lengths. The bovine yolk sac with about 9 cm on day 25 p. i. of pregnancy permanently decreased its total length during this study. Histologically, the yolk sac is composed of three cell layers: the mesothelium, the mesenchyme and the endoderm. In mesenchyme are found blood islets. In the endoderm are formed cells invaginations toward the mesenchyme originating small canaliculi. The ultrastructure of yolk cells presented many mitochondria, rough endoplasmic reticulum, vesicles, euchromatin and the presence of two nucleoli. Discussion: The real fi rst blood circulation in the bovine is attached with the development of yolk sac, differently from other membranes, such as the corium, that does not present evidence of vascularization by the age of 20-30 days. The erythroblasts found in the yolk sac are related to vasculogenesis and the process of differentiation of blood cells during the erythropoiesis. It could be observed on the histology of the yolk sac, in embryos of 30-50 days old, the presence of canaliculi and small folds of the epithelium. The canaliculi collapse is associated with the degeneration of the endoderm wall of the yolk sac. The organelles present in the endoderm cells of the yolk sac are associated with the function of protein metabolism and in the exchange of substances between the mesenchyme and the mesothelium. For these fi ndings, could be observed that the yolk sac epithelium is found active until the 50th day of gestation, and thereafter regresses. However, remnants of this membrane may be present until the 70th day. These features may represent a presence of an active chorionvitelline placenta in this period responsible for the maintenance of pregnancy whereas the chorioallantoic placenta is not defi nitively established.

Production of egg yolk antibody (IgY) against recombinant canine parvovirus VP2 protein.

Abstract: Background: Canine parvovirus-2 (CPV-2) disease is highly contagious and often fatal in canine with symptoms of hemorrhagic diarrhea and myocarditis. Three variants, CPV-2a, CPV-2b and CPV-2c, had emerged and replaced the CPV-2 in the past decades worldwide. Emergences of the new variants may increase the difficulty of CPV diagnosis and treatment. CPV capsid consists of 60 copies of a combination of viral coat proteins VP1 (82 kDa), VP2 (67 kDa) and VP3 (63.5 kDa), while about 90% of the capsid proteins are VP2 and most of the B cell epitopes are located on the VP2. IgY technology has been recognized as a promising alternative to generate large amount of qualified high specific antibody for use in immunodiagnostic and in immunotherapy with the advantages of relatively simple, noninvasive method and large-scale production over mammalian antibody. Furthermore, IgY antibody does not react with mammalian IgG nor binding to the rheumatoid factor, which may reduce the false positive results in immunoassays. Anti-VP2 IgY antibody has never been reported before, here we describe a method for the production of anti-VP2 IgY, which could be applied in the diagnosis and treatment for the CPV infection. Materials, Methods & Results: A CPV strain was isolated from a clinical sample. The VP2 ORF (Open reading frame, ORF) was amplified by PCR and inserted into the pMD18-T clone vector. The isolate was defined as CPV-2a (JN403045) subtype by sequencing. The VP2 ORF was inserted into pET-32a by T4 ligase and introduced into the E. coli Bal21. VP2 protein was produced by the induction of the E. coli Bal21 containing pET-32a-VP2 with isopropyl-β-D-1-thiogalactopyrannoside (IPTG). Expression of the recombinant VP2 protein (rVP2) fused with His-tag was analyzed by SDS-PAGE and detected by western blotting using anti-His monoclonal antibody. The rVP2 was purified by Ni+-affinity purification chromatography under denature condition and dialyzed against PBS. The concentration of the rVP2 was determined by Bradford method. After immunizing the chickens with rVP2, anti-VP2 IgY was isolated by PEG 6000 precipitation and analyzed by SDS-PAGE. The activity and specificity of the IgY antibody were analyzed by indirect ELISA and western blotting. SDS-PAGE analysis showed that the rVP2 fused with His-tag had a molecular of 85 kDa, accompanied with the low molecular fractions around 50 kDa and 40 kDa. The rVP2 could be recognized be the anti-His monoclonal antibody. The IgY antibody isolated by PEG 6000 method and analyzed by SDS-PAGE showed the IgY mainly contained two parts, 23 kDa and 67 kDa, which corresponded to light and heavy chain, respectively. In additional, some lower bands around 40 kDa were presented on the gel. The anti-VP2 IgY reached to 1:40960 after the fourth immunization. The anti-VP2-IgY could recognize the VP2 specially in Western blotting, while no reaction was seen with the low molecular. Discussion: The emergence of the low molecular proteins in 50 kDa and 40 kDa in the pellets of the bacterial lysates may be due to the degradation of the rVP2 by endogenous proteases in E. coli. The fact that IgY could recognize the entire VP2 fraction suggests the lost of the antigenic sites of the low molecular proteins.

In Vitro Evaluation of the Antiviral Potential of Guettarda angelica Against Animal Herpesviruses

Abstract: Background: The number of antiviral studies using plant extracts has increased in the last decades, and the results have shown that plants are potential sour...

Surgical Treatment of Corneal Large Dermoid in a Simmental Calf

Abstract: Background: Ocular dermoid is a skin or skin-like appendage usually arising on the limbus, conjunctivae, and cornea. It can be unilateral or bilateral and may be associated with other ocular manifestation or with other malformations. Hair from the lesions is predominantly responsible for the associated irritation resulting in chronic inflammation of the conjunctivae and cornea and may cause visual impairment. Ocular dermoids are rare in cattle, with the prevalence estimated between 0.002% and 0. 4%. Case: A one-day-old Simmental calf presented for bilateral haired masses on the eyes and tumor-like growth since birth. The calf was admitted to the University of Adnan Menderes, Faculty of Veterinary Medicine 1 days after initial examination. The calf weighed 35 kg, was in good bodily condition and no further abnormalities were detected on physical examination, complete blood count and serum chemistries. General anesthesia was induced and maintained with an intravenous detomidine-ketamine-midazolam combination. The corneal dermoids extended slightly beyond the inferonasal limbus and then merged with a second mass of lightly haired tissue within the inferonasal bulbar conjunctiva of both eyes. The dermoids were excised by superficial lamellar keratectomy that was extended into the inferonasal conjunctiva. The nasal masses were excised by electrocautery. All excised tissue was submitted for histopathology. Histopathology of the excised tissue confirmed bilateral corneoconjunctival dermoids with bilateral nasal hyperplastic tissue. Topical 1% atropine sulfate ointment was applied twice daily to both eyes for 5 days and bacitracin-neomycin-polymixin ophthalmic ointment applied twice daily to both eyes for 2 weeks postoperatively. Ocular and nasal healing was uneventful, with prominent corneoconjunctival granulation tissue 6 days postoperatively, which largely resolved by the 6-week follow-up examination. Bilateral large dermoids with nasal hyperplastic tissue have not been documented previously in Simmental cattle in Turkey. Discussion: The precise developmental mechanisms involved in the pathogenesis of ocular dermoids are not known. Metaplasia of mesenchyme (of primarily neural crest origin), resulting in abnormal differentiation of the surface ectoderm, is considered the most likely mechanism. Ocular dermoids may be associated with other congenital ocular or multiorgan abnormalities. The medial and lateral nasal processes, optic vesicle and first and second branchial arches are in close relationship on the lateral side of the embryonic head and might all be adversely influenced by the same stimulus. Ectopic lacrimal tissue may appear in combination with an ocular dermoid as in this case report and previously reported in cattle, or alone as an epibulbar choristoma. The ectopic lacrimal tissue in this calf may have represented misplacement of the nictitans gland or additional lacrimal tissue development. Superficial keratectomy is required to surgically excise a corneal dermoid although the depth of the dermoid within the cornea cannot be ascertained by ophthalmic examination until surgery is undertaken. In the case of large corneal dermoids, surgical excision should be performed early in the life of the patient to achieve optical improvement and allow functional development of the eye.

Evaluation of a Latex Agglutination Kit for Detecting Rotavirus in Piglets

Abstract: Background: The group A rotavirus is the most important ethiological agent that causes diarrhea in newborn humans and animals. They belong to the Reoviridae family and have a genome consisting of 11 segments of double-stranded RNA enclosed in a triple-layered capsid. Rotaviruses are classifi ed in six groups (A to G) based on the VP6 capsid protein or on the migration pattern of genomic segments in polyacrylamide gel. Groups A, B, and C were found either in humans or in animals, while groups D to G were found only in animals. The outer layer is formed by two proteins, VP7 and VP4, which elicit neutralizing antibody responses and form the basis of the current dual classifi cation system in G (VP7) and P (VP4) types. The early diagnostic of this viral infection in raising farms is crucial for preventing the dissemination of the disease among the animals. Latex agglutination tests (LAT) are quick and easily done, and require low fi nancial investments. Materials, Methods & Results: The present study evaluated the Rotavirus Latex® kit (Richmond Immunosystems Diagnostics) currently used with humans for detecting the presence of rotavirus antigen in diarrheic feces of piglets. In order to confi rm the results obtained with the LAT, samples were analyzed by EIA with the Ridascreen rotavirus® kit (R-Biopharm). The results were compared with the rotavirus determination test by polyacrylamide gel electrophoresis (PAGE). During March 2007 and March 2008, 328 excrement samples of diarrheic piglets were collected at a hog-raising farm in the state of Rio Grande do Sul, Brazil. Results showed that the rotavirus was present in 38 samples (11.6%) assayed with LAT, while PAGE detected the virus in 26 samples (7.9%). This means that 15 positive cases (4.6%) were not identifi ed by PAGE. This difference shows the higher sensitivity of LAT against PAGE. LAT showed 88.5% of sensitivity, 95% of specifi city, 60.5% of positive predictive value, 99% of negative predictive value and 94.5% of accuracy. Discussion: Latex agglutination is easy to perform in a short time and does not require expensive equipment or skilled personnel, and the reagents have long shelf lives. These factors make the LAT suitable and highly effi cient for use in a clinical laboratory and a small farm as a rapid screening test for piglet rotavirus. A test used for detecting the etiological agent of gastroenteritis should be fast, easy to use, and specifi c for anticipating the disease treatment, thus minimizing unnecessary expenses and establishing prophylactic measures to protect the entire fl ock. The most important factor in choosing the method is the number of samples that should be collected, the qualifi cation of the analysis laboratory, and the type of pathogen that has contaminated the fl ock. The diagnostic methods most easily applied are based on detecting viral particles or RNA from fecal samples. Most kits available in the market for detecting rotaviruses are designed for humans and are not adequate for use in veterinary diagnosis. Therefore, the results obtained herein presented high sensitivity and specifi city for LAT, showing to be a valuable tool for diagnosing rotaviruses in pig feces. This system used in clinical laboratories might also be used in intensive animal farming systems or by small-scale pig raisers in a cooperative system.

Detecção de Salmonella enterica e Listeria monocytogenes em carcaças suínas na etapa de pré-resfriamento

Abstract: Background: Salmonellosis is one of the most prevalent foodborne diseases worldwide. Although listeriosis has been quite less reported, it is considered a major public health hazard due to the severity of symptoms caused in humans. Previous studies demonstrated that the genus Listeria and Salmonella can be found infecting pigs in Brazil, however there are yet few reports about their isolation from carcasses after the slaughtering process. From this, the aims of this study were to investigate the presence of Salmonella enterica and Listeria monocytogenes on pre-chill pig carcasses and to evaluate the presence of fecal carriers at the lairage. Materials, Methods & Results: Two sampling events were conducted in each of three slaughterhouses located in the state of Santa Catarina, Brazil. In each sampling event, pen feces from pigs belonging to three slaughter batches originated from different farms were collected. Thereafter, swabs were taken on the surface (loin, jowls, belly and ham) of 42 carcasses belonging to the same pig batches sampled at the lairage. All samples were submitted to a protocol for isolation of S. enterica and L. monocytogenes. Moreover, coliforms were enumerated in the samples taken from the carcasses. From a total of 18 samples of pen feces, 83.3% (15/18) were positive for S. enterica. The genus Listeria was isolated from 66.7% (12/18) of pen feces samples, but only one isolate was confi rmed as L. monocytogenes. Among the 252 pre-chill carcasses sampled, S. enterica was isolated from 27.4% (69/252); and S. Typhimurium was the most frequent serovar identifi ed. On the other hand, L. monocytogenes was detected in 19.8% (50/252) of the carcasses. In slaughterhouse B, a signifi cantly higher frequency (P < 0.001) of L. monocytogenes isolation than in the other slaughterhouses was observed. S. enterica was signifi cantly more (P < 0.001) isolated than L. monocytogenes in the other two sampled slaughterhouses. The coliform mean counts found on carcass samples ranged from 1.25 x 100 to 8.25 x 104 . In slaughterhouse A, the coliform mean was signifi cantly lower (P < 0.001) than the mean of coliforms determined in the other slaughterhouses. Discussion: The signifi cantly higher frequency of L. monocytogenes isolation from carcasses sampled at the slaughterhouse B suggests that there were fl aws in its slaughter process. Nevertheless, the L. monocytogenes contamination on the carcasses may not have been originated from feces, since this Listeria specie was found in only one sample of pen feces. In spite of the fact that slaughterhouse A presented a signifi cantly lower mean of coliforms on the pre-chill carcasses, indicating that there was a better hygiene in the slaughtering process, the frequency of S. enterica on the carcasses was not different in comparison to those found in the other sampled slaughterhouses. This result may be related to the large number of fecal carriers detected at the lairage, which in turn may have led to an increase on the carcass contamination hazard throughout the slaughter line. In conclusion, S. enterica and L. monocytogenes can be isolated from pre-chill pig carcasses. Whereas the high number of Salmonella fecal carriers may have contributed to the frequency of contaminated carcasses, L. monocytogenes strains found on carcasses were probably not originated from feces.

Complications associated with ovariohysterectomy using nylon tie-rap as an hemostatic method.

Abstract: Background: Routine neutering of bitches is typically carried out by ovariohysterectomy (OVH) in Veterinary Medicine, seen by many as a simple procedure. During the conventional approach the points of attachment of the uterus and ovaries are double ligated with absorbable or non-absorbable sutures or even with titanium clamps. It has been described the use of non-surgical polyamide tie-raps as a hemostatic closure method by some authors but there are few papers describing the consequenses of such use. The aim of this study is to report four cases of surgical complications associated with nylon tie-raps use on OVHs on bitches referred to the Veterinary Teaching Hospitals of the Federal University of Rio Grande do Sul (HCV-UFRGS) and Passo Fundo University (HV-UPF), both at the State of Rio Grande do Sul. Cases: 1An eleven-month-old bitch was admitted at the HV-UPF nine days after OVH performed elsewhere, with omentum evisceration due to suture dehiscence. During emergency surgery both ovaries were found surrounded by nylon tie-raps and fi brous tissue adherence. Peritonitis signs were recognizable. Bilateral ovariectomy with nylon 2-0 was performed and the remaining uterus was removed. 2A fi ve-years-old bitch was referred to the HV-UPF ten days after OVH performed elsewhere, with apathy and suture dehiscence. Free liquid was identifi ed around the bladder and kidneys using the ultrasound exam. Intense hemorrhage from uterine vessels was found during celiotomy. A nylon tie-rap was found loose on the abdominal cavity and another couple were found at both ovaries surrounded by fi brous tissue adherence. Bilateral ovariectomy with nylon 2-0 was performed and a blood transfusion was necessary. 3A three-years-old bitch was admitted at the HCV-UFRGS six days after OVH performed at another institution, presenting suture dehiscence and evisceration. During emergency surgery, bladder, intestine and musculature necrosis due to the contact with sharp edged nylon tie-raps (prior placed as hemostatic devices), intense fi brous tissue adherence and peritonitis were identifi ed. 4A two-years-old bitch was referred to the HCV-UFRGS one year after OVH performed elsewhere, with a four-month persistent sinus tract on the right fl ank unsuccessfully treated by another veterinarian. Abdominal communication was suspected and therefore the patient was submitted to exploratory celiotomy. During surgery intense fi brous tissue adherence were found on both mesovarium. Stump granuloma and a sinus tract were also identifi ed in the right side. Sharped nylon tie-raps were found among the fi brous tissue and were then replaced by non-absorbable suture. Discussion: OVH complications such as suture dehiscence, evisceration, peritonitis and loosen tie-raps, all seen on cases 1, 2 and 3 are usual when there is trans-operatory contamination and can probably be associated with technique failure, as described by other authors. Self-trauma must be avoided throughout the use of Elizabethan collar, but this recommendation was not followed by the patients’ owners on any of the cases even though it had been prescribed by the veterinarian. Fibrous tissue and granulomas were found on all cases due to the tie-raps, what brings the question about its real effects to the animal organism. All the patients had satisfi able recovery after the emergency surgeries. There is urgent need of scientifi c investigations about the grave consequences of the inadequate use of poliamide tie-raps before assigning it as a hemostatic material.

Doxycycline-chloroquine combination for the treatment of canine monocytic ehrlichiosis

Abstract: Background: Ehrlichiosis in dogs is a vector borne disease caused by Ehrlichia canis, from the Anaplasmataceae family, capable of causing multisystemic disorders. Following an incubation period of 8-20 days, acute, subclinical and chronic forms of the disease may develop and affected dogs frequently showed hemorrhage, lymphadenopahty, splenomegaly, hepatomegaly, along with cardiac/renal disorders and myelosupression. Most of the untreated dogs spontaneously recover form the acute phase and enters the subclinical phase, in which some of them may develop chronic phase characterized by bone marrow aplasia. Ehrlichial organisms are commonly susceptible to tetracycline derivatives, and doxycycline is probably most commonly used for treatment of the infection. Doxycycline may be quite effective for clearing parasitemia in acute E. canis infection. Clinical recovery may be observed within 2-3 days, besides treatment should be continued for 3 weeks, as some cases may remain carriers even if short treatment protocole is administered. Most dogs infected with CME usually recover from the acute and subclinical phases when treated with doxycycline or other tetracyclines. Some dogs enter the chronic phase of the disease for which the prognosis is grave. The purpose of the present study was to report the presence of E. canis infection in dogs in Aydin, with a special reference to the efficacy of combined doxycycline and chloroquine therapy. Materials, Methods & Results: A propective study was carried out on dogs referred and 12 dogs were diagnosed as canine monocytic ehrlichiosis within traditional buffy coat smear within giemsa staining, Snap 4dx test and nested PCR applications. Data on rectal temperature, clinical haematology and serum biochemistry (involving vbALT, AST and BUN) were registered before and after treatment in both groups. A total of 6 dogs with a diagnosis of CME were enrolled in each group. Group CD (n = 6) received doxycycline (10 mg/kg perorally twice a day for 2 weeks) and chloroquine (2.5 mg/kg perorally twice a day for 2 weeks) and group D received solely doxycycline (10 mg/kg perorally twice a day for 2 weeks). The clinical haematology and biochemistry results of canine ehrlichiosis before (day 0) and after treatment (day 30) for both gorups were compared. Among relevant parameters of aforementioned groups, no statistically significant differences were detected (P > 0.05). Following treatment in both groups the clinical signs were dinished, body temperature of the dogs returned to physiological levels in both groups. Lymphadenopathy was regressed in 1 week, and 1 month later the clinical examination was repeated in which all dogs in combined treated group showed complete clinical recovery, whereas lymphadenopathy was stil evident in some of the dogs in chloroquine group. Discussion: Doxycycline has still been the first line-drug option for the treatment of acute canine monocytic ehrlichiosis, however for the subclinical and chronic phases of the diseases its effectiveness is controversial. Further research are warranted to investigate any adjuvant or combined therapy may be an alternatives. In this clinical trial combined doxycycline and chloroquine therapy were used for dogs with active Ehrlichia canis infection (PCR + and E. canis seroactive). Although there were no statistically significant hematological and serum biochemical differences among dogs treated with doxycycline and chloroquine in contrast to dogs treated alone with doxycycline, clinical recovery was impressive in dogs with combined treatment. These observations demonstrate that chloroquine may have helped hastening the relevant clinical signs of canine monocytic ehrlichiosis and clinical improvement.

Lobação pulmonar e distribuição brônquica do ouriço-cacheiro (Sphiggurus villosus)

Abstract: Background: The orange-spined hairy dwarf porcupine (Sphiggurus villosus) is a mammal found in Brazil, Venezuela, Guyanas and Bolivia. Its dorsum is yellowis...

Leishmaniose Visceral Canina: Dois casos autóctones no município de Florianópolis, estado de Santa Catarina

Abstract: Background: Brazil faces an expansion and urbanization of American visceral leishmaniasis with human and canine cases in several metropolises. This report describes two cases of autochthonous canine visceral leishmaniasis in the city of Florianopolis, state of Santa Catarina, classified as an area previously without disease transmission. Case: Five dogs from Canto dos Aracas, Lagoa da Conceicao, Florianopolis municipality, which had no history of displacement to other municipalities, were studied by collecting clinical specimens in an attempt to isolate the parasite and to confirm the infection by Leishmania. For sampling the animals were sedated and after local shaving, asepsis and anesthesia with lidocaine HCl 2%, two fragments of apparently healthy skin of the scapular region with the aid of a 3 mm punch were obtained. The two fragments were placed in sterile saline plus antibiotics and antifungal agents solution to attempt the isolation of the parasite in NNN culture medium plus Schneider’s medium. In addition to the collection of skin fragment, animals underwent bone marrow (BM) needle aspiration, obtained from the manubrium of the sternum, using a 20 mL syringe with 40x12 mm needle and aspiration biopsy of popliteal lymph nodes using Valeri citoaspirador 10 mL syringe and 25x8 mm needle, both performed after local shaving, asepsis and anesthesia with 2% lidocaine. The material collected from punctures biopsies was also seeded in an attempt of isolating the parasites. Among the five examined animals, in three cases promastigotes were isolated, two of which through the cultivation of skin fragments and one through a punctured sample of the lymph node. In two cases isoenzymatic characterization and identifying parasites such as Leishmania chagasi could be performed. Discussion: The LVC findings in Florianopolis, Santa Catarina generates great concern due to lack of knowledge of the actual spread of the disease, transmission dynamics in this area, knowledge of the vector, wild reservoirs and even in relation to the canine population exposed, for the first time, to Leishmania chagasi. These peculiar conditions may contribute to the onset of an abrupt expansion of the disease, differently from endemic regions. This fact can also be observed in many other cities in Brazil, mainly due to the development and expansion of road networks and the popularity of air travel, which increases the transit of people and their pets thereby increasing the risk of dispersion, not only of LVC, as of other infectious diseases to non-endemic areas. Another important point which causes dispersion of the LVC is related to some owners’ attempt to get around the main control method, which is euthanasia for positive dogs, and end up moving their animals to areas where there is no endemic canine serological survey, endangering the population of these areas. The use of some tools could avoid this type of dispersal, as the tighter control of the movement of dogs from endemic areas, implementation of mandatory serological diagnosis and mandatory reporting of positive canine cases. Moreover, educational measures to educate dog owners and veterinarians from the public and private sectors would also help in trying to minimize this problem. In this sense, it is extremely important that the entomological and epidemiological surveillance aimed at LVC are deployed and implemented in free areas, aiming at detecting the presence of vector and/or canine enzootic diseases before installing the anthropozoonotic cycle, allowing unprecedented action aimed at preventing the occurrence of disease in humans, or even prepare the health services and the population to fight the problem.

Spatial distribution of seropositive dogs to Leptospira spp. and evaluation of Leptospirosis risk factors using a decision tree

Abstract: Background: Leptospirosis is an important zoonotic disease associated with poor areas of urban settings of developing countries and early diagnosis and prompt treatment may prevent disease. Although rodents are reportedly considered the main reservoirs of leptospirosis, dogs may develop the disease, may become asymptomatic carriers and may be used as sentinels for disease epidemiology. The use of Geographical Information Systems (GIS) combined with spatial analysis techniques allows the mapping of the disease and the identifi cation and assessment of health risk factors. Besides the use of GIS and spatial analysis, the technique of data mining, decision tree, can provide a great potential to fi nd a pattern in the behavior of the variables that determine the occurrence of leptospirosis. The objective of the present study was to apply Geographical Information Systems and data prospection (decision tree) to evaluate the risk factors for canine leptospirosis in an area of Curitiba, PR. Materials, Methods & Results: The present study was performed on the Vila Pantanal, a urban poor community in the city of Curitiba. A total of 287 dog blood samples were randomly obtained house-by-house in a two-day sampling on January 2010. In addition, a questionnaire was applied to owners at the time of sampling. Geographical coordinates related to each household of tested dog were obtained using a Global Positioning System (GPS) for mapping the spatial distribution of reagent and non-reagent dogs to leptospirosis. For the decision tree, risk factors included results of microagglutination test (MAT) from the serum of dogs, previous disease on the household, contact with rats or other dogs, dog breed, outdoors access, feeding, trash around house or backyard, open sewer proximity and fl ooding. A total of 189 samples (about 2/3 of overall samples) were randomly selected for the training fi le and consequent decision rules. The remained 98 samples were used for the testing fi le. The seroprevalence showed a pattern of spatial distribution that involved all the Pantanal area, without agglomeration of reagent animals. In relation to data mining, from 189 samples used in decision tree, a total of 165 (87.3%) animal samples were correctly classifi ed, generating a Kappa index of 0.413. A total of 154 out of 159 (96.8%) samples were considered non-reagent and were correctly classifi ed and only 5/159 (3.2%) were wrongly identifi ed. On the other hand, only 11 (36.7%) reagent samples were correctly classifi ed, with 19 (63.3%) samples failing diagnosis. Discussion: The spatial distribution that involved all the Pantanal area showed that all the animals in the area are at risk of contamination by Leptospira spp. Although most samples had been classifi ed correctly by the decision tree, a degree of diffi culty of separability related to seropositive animals was observed, with only 36.7% of the samples classifi ed correctly. This can occur due to the fact of seronegative animals number is superior to the number of seropositive ones, taking the differences in the pattern of variable behavior. The data mining helped to evaluate the most important risk factors for leptospirosis in an urban poor community of Curitiba. The variables selected by decision tree refl ected the important factors about the existence of the disease (default of sewer, presence of rats and rubbish and dogs with free access to street). The analyses showed the multifactorial character of the epidemiology of canine leptospirosis.

A Hematologic and Electrophoretic Study in Puppies Vaccinated Against Canine Distemper Virus and Canine Parvovirus

Abstract: Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus. Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein fractions alpha-1 globulin (68%) and alpha-2 globulin (41.4%) at day 7. Moreover, a 5-fold increase in the fibrinogem concentration and in the number of eosinophils was observed at day 14. Thereafter, the platelet count decreased by 27.3% and the number of monocytes increased 5-fold at day 28. Discussion: Mild to moderate thrombocytopenia is often observed in dogs 3-5 days post-vaccination with live attenuated vaccines, mainly those against CDV and CPV. Besides the platelet damage caused by the CDV per se, infected animals showed secondary immune-mediated thrombocytopenia and decreased platelet production due to direct viral megakaryocyte infection. The increase in alpha-1 globulin may be related to the augment in the synthesis of alpha-1 antitrypsin, the main protein of the alpha-1 globulin region, in response to the vaccine-induced acute inflammatory process. The alpha-2 globulin region includes haptoglobin, alpha-2 macroglobulin and ceruloplasmin, and the increase observed in this fraction suggested that both haptoglobin and ceruloplasmin levels were augmented, following acute inflammatory response pattern. Fibrinogen is a soluble plasma glycoprotein that is converted by thrombin into fibrin during blood clotting. Despite the increase in fibrinogen concentration be the best indicator of inflammation in large animals, the hyperfibrinogenemia observed suggests that the inflammatory process was adequate to stimulate synthesis of this acute phase protein (P < 0,05). Absence of lymphocytosis observed at days 49, 56 and 63 associated to the progressive increase of the gamma globulin fraction, although not statistically significant, suggested an augment of B lymphocytes. The eosinophilia was observed in highlighting the presence of inflammation. Moreover, an increase in monocyte count indicating the presence of subacute or chronic inflammation after the second dose of the vaccine.

Use of FTA cards for the transport of DNA samples of Salmonella spp. from poultry products from Southern Brazil.

Abstract: Background: The contamination of products with Salmonella is a major threat to the poultry industry because the possible transmission to humans and animals can produce a huge negative impact. The diversity of Salmonella enterica serotypes complicates the diagnostic systems and the transport of live cultures to the diagnostic labs may represent a biohazard. Current methods for serotyping using antibodies do not work well for many Salmonella serotypes and reagents are not often available. For these reasons, methods that assign serotype by the analysis of DNA are preferred. One step that is currently in development is streamlining methods for DNA submission to the laboratories for sequencing. For this purpose, we investigated fi lter papers commercially available (Flinders Technology Associates FTA) to ship DNA samples. Filter papers are impregnated with a chemical formulation that lyses cells, immobilizes DNA, and protects it from degradation. The objective of this study was to assess the feasibility of the FTA cards for transporting Salmonella DNA samples in order to reduce biohazards and if they would yield enough DNA in quantity and quality for molecular analyses. Material, Methods & Results: In this study 156 samples of Salmonella enterica serotypes Enteritidis, Heidelberg, Hadar, Gallinarum, Typhimurium, Agona and Pullorum were isolated from poultry products and environments in southern Brazil. Samples were stored in the Avian Diagnostic and Research Center of the Federal University of Rio Grande do Sul. Following instructions for spotting cards with cell cultures at a density that visually matched a McFarland Turbidity Standard 0,5; they were shipped to the Agriculture Research Service of the United States Department of Agriculture (USDA-ARS, Athens, GA-USA), using FTA cards. Upon the reception of the cards, safety testing was performed by transferring one disk from each sample into 10 mL of brain heart infusion (BHI) tubes and incubated at 37°C for 24 h. The BHI tube that showed turbidity after incubation was transferred to brilliant green (BG) agar and incubated at 37°C for 24 h to 48 h. If colonies were obtained in BG, biochemical analyses were performed by using the Enterotube method. Only one sample (S. Enteritidis) showed turbidity in BHI, but any bacterial growth was observed in the BG agar. The average DNA concentration, as measured by spectrophotometry, was 42,32 (± 9,84) ng/μL and the average 280/260 ratio was 1,9 (± 0,09). All the analyzed samples were negative for live cultures of Salmonella and the DNA obtained was suitable for molecular testing. Discussion: FTA cards can be used to transport DNA samples from pathogenic bacteria, reducing biohazards associated with shipping live cultures. The possibility of shipping DNA, in an economic and safe way, for testing samples at the laboratories facilitates the identifi cation of Salmonella enterica serotypes that are circulating in the environment of poultry. Turbidity in BHI tubes that did not result in colonies on agar media may be caused by the presence of other contaminants such as environmental saprophytic microorganisms that may occurred during the process of handling the cards. DNA samples of Salmonella enterica shipped from Brazil to the United States for this set of isolates did not show bacterial growth. Thus the FTA cards provided safe and effective inactivation of the pathogen, and the DNA obtained from the cards were adequate for downstream analyses.

Uso dos cartões FTA para o transporte de amostras de DNA de Salmonella spp. isoladas de produtos avícolas do Sul do Brasil

Abstract: Background: The contamination of products with Salmonella is a major threat to the poultry industry because the possible transmission to humans and animals can produce a huge negative impact. The diversity of Salmonella enterica serotypes complicates the diagnostic systems and the transport of live cultures to the diagnostic labs may represent a biohazard. Current methods for serotyping using antibodies do not work well for many Salmonella serotypes and reagents are not often available. For these reasons, methods that assign serotype by the analysis of DNA are preferred. One step that is currently in development is streamlining methods for DNA submission to the laboratories for sequencing. For this purpose, we investigated fi lter papers commercially available (Flinders Technology Associates - FTA) to ship DNA samples. Filter papers are impregnated with a chemical formulation that lyses cells, immobilizes DNA, and protects it from degradation. The objective of this study was to assess the feasibility of the FTA cards for transporting Salmonella DNA samples in order to reduce biohazards and if they would yield enough DNA in quantity and quality for molecular analyses. Material, Methods & Results: In this study 156 samples of Salmonella enterica serotypes Enteritidis, Heidelberg, Hadar, Gallinarum, Typhimurium, Agona and Pullorum were isolated from poultry products and environments in southern Brazil. Samples were stored in the Avian Diagnostic and Research Center of the Federal University of Rio Grande do Sul. Following instructions for spotting cards with cell cultures at a density that visually matched a McFarland Turbidity Standard 0,5; they were shipped to the Agriculture Research Service of the United States Department of Agriculture (USDA-ARS, Athens, GA-USA), using FTA cards. Upon the reception of the cards, safety testing was performed by transferring one disk from each sample into 10 mL of brain heart infusion (BHI) tubes and incubated at 37°C for 24 h. The BHI tube that showed turbidity after incubation was transferred to brilliant green (BG) agar and incubated at 37°C for 24 h to 48 h. If colonies were obtained in BG, biochemical analyses were performed by using the Enterotube method. Only one sample (S. Enteritidis) showed turbidity in BHI, but any bacterial growth was observed in the BG agar. The average DNA concentration, as measured by spectrophotometry, was 42,32 (± 9,84) ng/µL and the average 280/260 ratio was 1,9 (± 0,09). All the analyzed samples were negative for live cultures of Salmonella and the DNA obtained was suitable for molecular testing. Discussion: FTA cards can be used to transport DNA samples from pathogenic bacteria, reducing biohazards associated with shipping live cultures. The possibility of shipping DNA, in an economic and safe way, for testing samples at the laboratories facilitates the identifi cation of Salmonella enterica serotypes that are circulating in the environment of poultry. Turbidity in BHI tubes that did not result in colonies on agar media may be caused by the presence of other contaminants such as environmental saprophytic microorganisms that may occurred during the process of handling the cards. DNA samples of Salmonella enterica shipped from Brazil to the United States for this set of isolates did not show bacterial growth. Thus the FTA cards provided safe and effective inactivation of the pathogen, and the DNA obtained from the cards were adequate for downstream analyses.

Condrodisplasia tipo Dexter em fetos bovinos abortados

Abstract: Background: Chondrodysplasia is a congenital anomaly related to defects of the genes that control the chondrogenesis. This anomaly is described in many breeds of cattle and consanguineous matings increases the likelihood of developing defective traits. There are three distinct syndromes known: Dexter, Telemark and Brachycephalic type. The chondrodysplasia Dexter type syndrome is associated to an incompletely dominant gene, which occurs in Dexter and Holstein breeds more frequently, yet Charolais and Jersey can also be affected. There are three recognized phenotypes in this form of disease: severe achondroplasia, with abortion before the seventh month of gestation (monster Dexter), when related to dominant homozygous; chondrodisplasia with limbs shortening, when heterozygous; and normal animals, when recessive homozygous. The objective of this paper is to describe Dexter chondrodysplasia observed in two bovine fetuses examined by the Setor de Patologia Veterinaria from Universidade Federal do Rio Grande do Sul (SPV-UFRGS). Case: The necropsy of two aborted bovine calves was performed by the SPV-UFRGS. The Fetus 1 did not have data, neither clinical history. The Fetus 2, a Jersey breed calf, was in the last third of gestation and came from a property with Jersey and mini-Jersey herd. Natural mating was used for reproduction, with only one bull. Macroscopic alterations observed in both fetuses were characterized by shortened limbs, rounded and disproportionate skull, short snout, undershot jaw, tongue protrusion and abdominal hernia with eventration. In the second fetus was also observed spina bifi da and palatoschisis. Microscopically, in both fetuses were observed extensive areas of immature cartilage, chondrocytes of the femur’s bone tissue were not organized in recognizable growth plates, the metaphysis was markedly shortened and consisted in short bone trabeculae with occasional cartilage isles. Immunohistochemistry (IHC) to detect bovine viral diarrhea virus (BVDV) was negative. Discussion: The macroscopic and microscopic alterations in both fetuses were compatible with chondrodysplasia “monster Dexter” type. Such alterations, associated with dominant homozygotes, are due to defective endochondral ossifi cation with little growth in the length of long bones, meanwhile intramembranous ossifi cation beneath the periosteum keeps growing and contributing to the abnormal volume and growth of the bones. Such difference is clearly observed when the size of the head is compared with the rest of the body. As the skull bones depends basically on intramembranous ossifi cation, they are generally bigger than the rest of the body. The negative result of IHC to BVDV shows that this agent, known as the cause of congenital malformation did not have association with the anomalies observed. According to other researchers, this low frequency of viral agents as responsible for congenital anomalies was observed previously in cattle from the south of Brazil. Although Dexter chondrodysplasia can be considered of occasional occurrence, it can be responsible for considerable damage. As an abnormality with hereditary nature and consanguinity is the main reason to its occurrence, prevention is the main method to be adopted by the producer to avoid bigger loses.

Complicações associadas à ovariosalpingohisterectomia eletiva realizada com abraçadeira de náilon como método de hemostasia

Abstract: Cases: 1- An eleven-month-old bitch was admitted at the HV-UPF nine days after OVH performed elsewhere, with omentum evisceration due to suture dehiscence. During emergency surgery both ovaries were found surrounded by nylon tie-raps and fi brous tissue adherence. Peritonitis signs were recognizable. Bilateral ovariectomy with nylon 2-0 was performed and the remaining uterus was removed. 2- A five-years-old bitch was referred to the HV-UPF ten days after OVH performed elsewhere, with apathy and suture dehiscence. Free liquid was identified around the bladder and kidneys using the ultrasound exam. Intense hemorrhage from uterine vessels was found during celiotomy. A nylon tie-rap was found loose on the abdominal cavity and another couple were found at both ovaries surrounded by fibrous tissue adherence. Bilateral ovariectomy with nylon 2-0 was performed and a blood transfusion was necessary. 3- A three-years-old bitch was admitted at the HCV-UFRGS six days after OVH performed at another institution, presenting suture dehiscence and evisceration. During emergency surgery, bladder, intestine and musculature necrosis due to the contact with sharp edged nylon tie-raps (prior placed as hemostatic devices), intense fibrous tissue adherence and peritonitis were identified. 4- A two-years-old bitch was referred to the HCV-UFRGS one year after OVH performed elsewhere, with a four-month persistent sinus tract on the right flank unsuccessfully treated by another veterinarian. Abdominal communication was suspected and therefore the patient was submitted to exploratory celiotomy. During surgery intense fibrous tissue adherence were found on both mesovarium. Stump granuloma and a sinus tract were also identified in the right side. Sharped nylon tie-raps were found among the fibrous tissue and were then replaced by non-absorbable suture. Discussion: OVH complications such as suture dehiscence, evisceration, peritonitis and loosen tie-raps, all seen on cases 1, 2 and 3 are usual when there is trans-operatory contamination and can probably be associated with technique failure, as described by other authors. Self-trauma must be avoided throughout the use of Elizabethan collar, but this recommendation was not followed by the patients’ owners on any of the cases even though it had been prescribed by the veterinarian. Fibrous tissue and granulomas were found on all cases due to the tie-raps, what brings the question about its real effects to the animal organism. All the patients had satisfi able recovery after the emergency surgeries. There is urgent need of scientifi c investigations about the grave consequences of the inadequate use of poliamide tie-raps before assigning it as a hemostatic material.

Desvio facial (wry nose) em um equino adulto

Abstract: Background: Wry Nose (Campylorrhinus lateralis) is a congenital shortening and deviation of the maxillae, premaxillae, nasal bones, vomer, and nasal septum that affects mainly foals. Intrauterine positioning as an etiology of this deformity is highly unlikely. Clinically, varying degree of premaxillar deviation is noticeable upon inspection of the foal’s head. The deviation usually results in malocclusion, with no contact between the incisors of the mandible and maxilla. Some foals experience breathing diffi culties or have increased respiratory sounds. This condition will not self-correct, and a decision should be made to treat the foal using reconstructive techniques or to euthanize it to prevent it from suffering. The objective of this work was to describe a case of wry nose in an adult male equine. Case: A 4-year-old, 300 kg, male SRD equine was submitted for necropsy. There was no history of respiratory problems and trouble with mastication or prehension of food. Three days before death the horse showed clinical sings characterized by no ingestion of food and water, changing behavior and locomotion, weakness, and not respond to calls, seemingly oblivious to the environment evolving into permanent recumbency and death after about 24 h of the decline. The horse was treated by a veterinarian who suspected of leucoencephalomalacea. The animal died 24 h after her condition worsened. Macroscopically, the main lesion observed was in the nose. The nasal septum was deviated to the left with the sigmoid appearance and a dorsal hump on the right side of the nasal bone. The radiographs of dorsoventral view of the jaws showing premaxillae deviated approximately 90 degrees to the left. The left premaxillae was shorter than the right premaxilla, thus creating a lesser and greater curvature of the premaxilla. Macroscopic and Microscopic lesions in the brain were not observed discarding the possibility of leucoencephalomalacea and other neurological diseases of infectious origin. Discussion: Wry nose is a relatively rare disease and have being reported in foals. In our study, it was not possible to determine the cause of this abnormality, but a computed tomography study has been associated this deformity with an underdeveloped pregnant uterus during the second half of pregnancy witch the narrow tip of the uterine horns provides only limited space for the developing cranial half of the fetus. Although the disease have been reported in young animals in which the surgical procedure in moderate to severe cases it is recommended, in the present case, the equine was four years old indicating that there is a possibility of the animal to reach adulthood, but with their commitment to quality of life. Several diseases, such as cystic degeneration, malformation of normal tissue, abscesses, fractures and neoplasms may cause thickening, malformation or deviation of the nasal septum in horse producing similar clinical sings. In the present case, the macroscopic lesions and radiography exam were useful to determinate the exact site, extent and nature of the involvement of the lesion. Despite the owner to report a clinical picture indicative of neurological disease, histological analysis of the brain ruled out this possibility. These fi ndings indicate that wry nose disease should be kept in mind when investigating causes of deviation of the nasal septum in equines.

Esporotricose zoonótica: procedimentos de biossegurança

Abstract: Background: Sporotrichosis is a fungal infection caused by Sporothrix schenckii species complex, which is distributed worldwide, especially in regions of tropical and subtropical climates. It can affect both humans and a great variety of animals, among which, the domestic dog and cat. Today is considered the subcutaneous mycosis more frequent in Brazil, due to the progressive increase of zoonotic transmission of the disease in the metropolitan region of Rio de Janeiro. Review: In the endemic region of zoonotic sporotrichosis in Rio de Janeiro,veterinarians, their assistants and owners of cats with sporotrichosis are considered risk groups for acquiring the infection. Of the human cases of sporotrichosis in this endemic region that were accompanied by the Clinical Research Institute Evandro Chagas (IPEC / Fiocruz) in the period from 1998 to 2001, 5% of the patients diagnosed for the disease were veterinarians, demonstrating the occurrence of the infection in this occupational group. Biosafety is defi ned as a condition of security achieved by a set of measures aimed at the prevention, control, reduction or elimination of risks involved in activities that could endanger the health of humans, animals, plants and the environment. However, small animal veterinary pratictioners from the endemic region of Rio de Janeiro usually do not follow biosafety measures, which increases the risk of acquisition of sporotrichosis. In most cases, the infection results from traumatic inoculation of the fungus in skin and in humans, clinical forms may be: fi xed or localized cutaneous, lymphatic-cutaneous, spread-cutaneous, mucosal and extra-cutaneous or systemic. In the endemic form of this disease that occurs in Rio de Janeiro, the cat is seen as a source of infection. The zoonotic potential of the cat is characterized by the abundance of yeasts found in their skin lesions and proximity with humans. Cats acquire the disease after fi ghts with other infected cats and the skin lesions more frequently found are nodules and ulcers, covered or not by crusts, which can progress to necrosis with exposure of bones and muscles. The presence of respiratory signs, especially sneezing, with or without lesions in the mucosal and in the nasal region is common. The azole itraconazole is considered the drug of choice for humans and cats. However, unlike humans, the treatment is considered diffi cult in cats. The defi nitive diagnosis of sporotrichosis is obtained by isolation of Sporothrix in culture. Nonetheless, in cats, due to the great quantity of yeasts in their lesions, cytopathological test is strongly indicated in the presumptive diagnosis because of the speed in processing, low cost and no requirement of sophisticated technical training or complex laboratory structure. Discussion: A zoonotic form of sporotrichosis has become endemic in the metropolitan region of Rio de Janeiro and the veterinarians are a high risk group for acquiring the disease as well as have a very important role in the application of measures for the prevention and control of sporotrichosis. Therefore, in this review, specifi c biosafety procedures to reduce risks during the handling of cats with suspected sporotrichosis by veterinarians, technicians, caretakers and owners of cats were described. The topics aproached were: clinical care of the cat (where were appointed the recommended personal protective equipment, animal restraint and good practices), decontamination of the environment, equipment and items used in the cat care and management of waste. Aspects related to the fungus and the disease itself were also discussed.

Efeito do mês de parição na taxa de gestação subsequente e no peso ao desmame dos bezerros de vacas Nelore

Abstract: Background: The concentration of births in the most appropriate time (months) of the year results in uniform lots of calves and allows the adoption of differ...

Parasitismo pelo ácaro de sistema respiratório Sternostoma tracheacolum em um canário belga (Serinus canaria domestica)

Abstract: Background: The nasal mites of the Rhinonyssidae family include the highest diversity of mandatory hematophagous endoparasites of the respiratory system of birds. In general, these mites cause no significant lesions, except Sternostoma tracheacolum that invades the trachea, lungs and air sacs and may compromise breathing in birds. This study reports the death of a Belgian canary (Serinus canaria domestica) due to the parasitism by S. tracheacolum with an aggravating and concurrent pododermatitis infection. Case: A Belgian canary from a commercial bird flock, in which a high mortality rate had lately been recorded, was submitted to necropsy. Multiple blackened foci (1.0 mm in diameter) present in the trachea and the air sacs were subsequently identified as Sternostoma tracheacolum mites. Microscopically, there were diffuse and moderate infiltrates of lymphocytes, plasma cells, macrophages and heterophils within the tracheal submucosa and areas of fibrosis and osseous metaplasia of the tracheal cartilage. In the air sacs, there were multiple yellowish-brown foci of caseous necrosis, which were surrounded by inflammatory infiltrate of macrophages and multinucleated giant cells. In the lungs, there were foci of caseous necrosis surrounded by macrophages. In the tracheal lumen, mites sections were observed. Clusters of nucleated erythrocytes and brown granular pigment were seen within the longitudinal section of the mites. Prussian blue (Perls) special staining evidenced no hemosiderin content in the mite’s tissues, in which no melanin could be detected, as well. Discussion: The parasitism by S. tracheacolum in canaries was first described in South Africa. In such cases, the mite induced clinical respiratory disease and killed the birds. Subsequently, reports of parasitism by these mites included several additional bird species, of which, Cockatiel (Nymphicus hollandicus) has been the most frequently affected and there are only a few reports of the condition in canaries. Clinically, hosts have difficulty breathing, exercise intolerance, coughing and sneezing. Depending on the severity, infection can persist for months or lead to death by suffocation or weakness. Clinical signs and death are related to the appearance of the lesions induced by the mites. Mite eggs are deposited in the lungs and the larvae hatch soon after oviposition. After the first meal, the female protonymph migrates to the posterior parabronchia, while the male protonymph remains in the lungs, where it completes the development. The adult mites full of eggs tend to occupy the parabronchia, the syrinx and the trachea. The negative results on Prussian blue and melanin bleach methods indicate that the dark pigment present within the tissues from the mites is neither hemosiderin nor melanin, respectively. In parasitism due to Plasmodium sp. (malaria), hemozoin, an iron-containing pigment associated with erythrocytic changes induced by the protozoan, has not been detectable by the Prussian blue staining. It is suggested that the dark color observed in the mites in this study could be associated with a similar pigment. The clinical respiratory diseases in birds are generally nonspecific. The paper focuses on the importance of the confirmation of the diagnosis by the necropsy and histopathological examinations, as well as on the characterization of this respiratory parasitic disease of birds.