Showing papers in "Veterinary Research in 2003"

Production effects related to mastitis and mastitis economics in dairy cattle herds.

Abstract: Mastitis is the most prevalent production disease in dairy herds world-wide and is responsible for several production effects. Milk yield and composition can be affected by a more or less severe short-term depression and, in case of no cure, by a long-acting effect, and, sometimes, an overlapping effect to the next lactation. Summary values in the literature for losses of milk production were proposed at 375 kg for a clinical case (5% at the lactation level) and at 0.5 kg per 2-fold increase of crude SCC of a cow. Due to the withdrawal period after treatment, composition changes in milk can almost be neglected in economic calculations. Lethality rate for clinical mastitis is very low on the average, while anticipated culling occurs more frequently after clinical and subclinical mastitis (relative risk between 1.5 and 5.0). The economics of mastitis needs to be addressed at the farm level and, per se, depends on local and regional epidemiological, managerial and economic conditions. To assess the direct economic impact of mastitis, costs (i.e. extra resource use) and losses (i.e. reduced revenues) have to be aggregated. To support decision making for udder health control, it is necessary to use a marginal approach, based on the comparison of the losses avoided and the additional costs of modified plans, compared to the existing ones.

Monitoring udder health and milk quality using somatic cell counts.

Abstract: In this article the use of somatic cell counts for monitoring udder health and milk quality is discussed. Somatic cell count dynamics at quarter, cow, herd and population level are discussed and illustrated with examples. Quarter and cow somatic cell counts directly represent the inflammatory status of the mammary gland. Herd and population somatic cell count are related to the inflammatory process in individual cows but much more reflect the udder health status of the herd and the quality of the raw milk in the herd and the population. Application of monitoring tools in herd health management are illustrated using a case study. Understanding infection dynamics requires precise longitudinal data. Monitoring tools are required to find the areas of risk in the herd. It is inevitable that more complete udder health programs and monitoring systems are to be developed and implemented. These programs are necessarily dynamic and complex. Implementation of complete udder health programs should be accompanied by research efforts to further fine-tune these complete udder health control and monitoring programs.

Mastitis of dairy small ruminants

Abstract: Staphylococci are the main aetiological agents of small ruminants intramammary infections (IMI), the more frequent isolates being S. aureus in clinical cases and coagulase negative species in subclinical IMI. The clinical IMI, whose annual incidence is usually lower than 5%, mainly occur at the beginning of machine milking and during the first third of lactation. These features constitute small ruminant peculiarities compared to dairy cattle. Small ruminant mastitis is generally a chronic and contagious infection: the primary sources are mammary and cutaneous carriages, and spreading mainly occurs during milking. Somatic cell counts (SCC) represent a valuable tool for prevalence assessment and screening, but predictive values are better in ewes than in goats. Prevention is most often based on milking machine management, sanitation and annual control, and milking technique optimisation. Elimination mainly relies on culling animals exhibiting clinical, chronic and recurrent IMI, and on drying-off intramammary antibiotherapy; this treatment allows a good efficacy and may be used selectively by targeting infected udders only. Heritability values for lactation mean SCC scores are between 0.11 and 0.15. Effective inclusion of ewe's mastitis resistance in the breeding goal has recently been implemented in France following experimental and large scale estimations of genetic parameters for SCC scores.

Severity of E. coli mastitis is mainly determined by cow factors

Abstract: Intramammary infections of dairy cows with Gram-positive bacteria such as Staphylococcus aureus (major cause of mastitis) have received a lot of attention because of their major economic impact on the dairy farm through production losses induced by an increase in somatic cell count. Management strategies, including greater awareness for efficient milking and hygienic measures, have limited the spread of Gram-positive bacteria and resulted in a significant decrease of proportion of S. aureus isolates and subclinical mastitis worldwide. Other organisms such as coliform subspecies and Streptococcus uberis, both environmental bacteria that cause clinical mastitis, have received less attention. Escherichia coli causes inflammation of the mammary gland in dairy cows around parturition and during early lactation with striking local and sometimes severe systemic clinical symptoms. This disease affects many high producing cows in dairy herds and may cause several cases of death per year in the most severe cases. It is well known that bacterial, cow and environmental factors are interdependent and influence mastitis susceptibility. Many studies, executed during the last decade, indicate that the severity of E. coli mastitis is mainly determined by cow factors rather than by E. coli pathogenicity. During E. coli mastitis, the host defense status is a cardinal factor determining the outcome of the disease. Today, we know that the neutrophil is a key factor in the cows' defense against intramammary infection with E. coli. Effective elimination of the pathogen by neutrophils is important for the resolution of infection and the outcome of E. coli mastitis. This review is a compilation of some major findings over the last 15 years concerning mainly host factors that modulate and influence neutrophil function and the mammary inflammatory reaction. The individual chapters address: virulence factors of E. coli strains, how neutrophils kill E. coli, connection between endotoxins, tumor necrosis factor-alpha and nitric oxide, severity classification of E. coli mastitis, lifespan of neutrophils, host factors that influence severity, tissue damage and production loss.

The bovine neutrophil: Structure and function in blood and milk.

Abstract: Migration of polymorphonuclear neutrophil leukocytes (PMN) into the mammary gland provide the first line of defense against invading mastitis pathogens. Bacteria release potent toxins that activate white blood cells and epithelial cells in the mammary gland to secrete cytokines that recruit PMN that function as phagocytes at the site of infection. While freshly migrated PMN are active phagocytes, continued exposure of PMN to inhibitory factors in milk such as fat globules and casein, leads to altered PMN morphology and reduced phagocytosis. In the course of phagocytosing and destroying invading pathogens, PMN release chemicals that not only kill the pathogens but that also cause injury to the delicate lining of the mammary gland. This will result in permanent scarring and reduced numbers of milk secretory cells. The life span of PMN is limited by the onset of apoptosis. To minimize damage to mammary tissue, PMN undergo a specialized process of programmed cell death known as apoptosis. Macrophages quickly engulf and phagocytose apoptotic PMN, thereby minimizing the release of PMN granular contents that are damaging to tissue. The PMN possess an array of cell surface receptors that allow them to adhere and migrate through endothelium and to recognize and phagocytose bacteria. One receptor found on phagocytes that is receiving considerable attention in the control of infections by Gram-negative bacteria is CD14. Binding of lipopolysaccharide (LPS) to membrane bound CD14 causes release of tumor necrosis factor-alpha and sepsis. Binding of LPS to soluble CD14 shed from CD14-bearing cells results in neutralization of LPS and rapid recruitment of PMN to the site of infection. Recent advances in the fields of genomics and proteomics should greatly enhance our understanding of the PMN role in controlling intramammary infections in ruminants. Further, manipulation of PMN, through either recombinant proteins such as soluble CD14 that enhance PMN response or agents that mediate PMN apoptosis, may serve as novel therapeutics for the treatment of mastitis.

Indicators of inflammation in the diagnosis of mastitis

Abstract: Mastitis affects the quality of milk and is a potential health risk for the other cows. In a well managed dairy herd, in addition to clinical mastitis, subclinical mastitis should be efficiently detected. Bacteriological sampling is not feasible as a routine test to identify subclinical mastitis, and indirect tests of mastitis are more suitable for selecting cows with intramammary infections for subsequent bacteriological sampling. Mastitis affects the composition of milk, and the degree of changes depends on the infecting agent and the inflammatory response. Indicators of inflammation in the milk which can be determined using rapid, reliable and easy routine techniques, can be used for the early detection of mastitis. The measuring of the somatic cell count in milk is the standard method, but the analysis technique is problematic for routine use in herds. The most promising parameters for monitoring subclinical mastitis are milk N-acetyl-beta-D-glucosaminidase activity, lactose, and electrical conductivity along with some other indicators such as optical and milk flow measurements, preferably with an inter-quarter evaluation included in the test. Acute phase proteins, haptoglobin and serum amyloid A, are also potential candidates for mastitis monitoring. New mastitis detection systems which can be adapted into on-line use are urgently needed, since dairy units are growing bigger and automatic milking systems are being taken into use.

Genetics of resistance to mastitis in dairy cattle.

Abstract: Genetic variability of mastitis resistance is well established in dairy cattle. Many studies focused on polygenic variation of the trait, by estimating heritabilities and genetic correlation among phenotypic traits related to mastitis such as somatic cell counts and clinical cases. The role of Major Histocompatibility Complex in the susceptibility or resistance to intrammamary infection is also well documented. Finally, development from molecular genome mapping led to accumulating information of quantitative trait loci (QTL) related to mastitis resistance and better understanding of the genetic determinism of the trait. From economic and genetic analyses, and according to welfare and food safety considerations and to breeders and consumer's concern, there is more and more evidence that mastitis should be included in breeding objective of dairy cattle breeds. Many countries have implemented selection for mastitis resistance based on linear decrease of somatic cell counts. Given biological questioning, potential unfavourable consequences for very low cell counts cows are regularly investigated. Improvement of selection accuracy for mastitis resistance is ongoing and includes: advances in modelling, optimal combination of mastitis related traits and associated predictors, such as udder morphology, definition of global breeding objective including production and functional traits, and inclusion of molecular information that is now available from QTL experiments.

Health hazards for terrestrial vertebrates from toxic cyanobacteria in surface water ecosystems

Abstract: Toxigenic cyanobacteria are photosynthetic prokaryotes that are most often recognized in marine and freshwater systems, such as lakes, ponds, rivers, and estuaries. When environmental conditions (such as light, nutrients, water column stability, etc.) are suitable for their growth, cyanobacteria may proliferate and form toxic blooms in the upper, sunlit layers. The biology and ecology of cyanobacteria have been extensively studied throughout the world during the last two decades, but we still know little about the factors and processes involved in regulating toxin production for many cyanobacterial species. In this minireview, we discuss these microorganisms, and more especially the toxins they produce, as a potential and important health risk for wild and domestic animals.

Metabolic changes in early lactation and impaired reproductive performance in dairy cows.

Abstract: This review addresses the suggestion that the decline in dairy reproductive performance, as increasingly observed these days, may be due to a hampered process of metabolic adaptation in early lactating cows. In our opinion, adaptation to the negative energy balance is a gradual process. Because almost all cows do adapt in the long run, it is not possible to classify animals as adapted or non-adapted. The use of risk factors is more appropriate in this case and is discussed in this review. Among them are the body condition score and its derivatives, feed intake, the calculated negative energy balance, and metabolic parameters like the plasma concentration of insulin or the triacylglycerol content in the liver. Moreover, factors that play a role in the link between declined reproductive performance and the metabolic situation of the cow during the early lactating period are discussed. Among these are insulin, insulin-like growth factors, leptin, neuropeptide Y, non-esterified fatty acids, thyroid hormones, urea, and ammonia.

Experimental Coxiella burnetii infection in pregnant goats: excretion routes

Abstract: Q fever is a widespread zoonosis caused by Coxiella burnetii. Infected animals, shedding bacteria by different routes, constitute contamination sources for humans and the environment. To study Coxiella excretion, pregnant goats were inoculated by the subcutaneous route in a site localized just in front of the shoulder at 90 days of gestation with 3 doses of bacteria (10(8), 10(6) or 10(4) i.d.). All the goats aborted whatever the dose used. Coxiella were found by PCR and immunofluorescence tests in all placentas and in several organs of at least one fetus per goat. At abortion, all the goats excreted bacteria in vaginal discharges up to 14 days and in milk samples up to 52 days. A few goats excreted Coxiella in their feces before abortion, and all goats, excreted bacteria in their feces after abortion. Antibody titers against Coxiella increased from 21 days post inoculation to the end of the experiment. For a Q fever diagnostic, detection by PCR and immunofluorescence tests of Coxiella in parturition products and vaginal secretions at abortion should be preferred to serological tests.

Effects of condensed tannins on established populations and on incoming larvae of Trichostrongylus colubriformis and Teladorsagia circumcincta in goats.

Abstract: The use of tanniferous plants or tannins represents one alternative approach to the control of gastrointestinal parasites in ruminants but most data have been obtained in sheep. The current study was therefore performed in goats with two objectives: firstly, to investigate the effects of condensed tannins (CT) on adult populations of Trichostrongylus colubriformis and Teladorsagia circumcincta; secondly, to examine their effects on the establishment of infective larvae of these two species. In experiment 1, two groups of kids were infected with 6 000 L3 of T. colubriformis and 6 000 L3 of T. circumcincta. After 7 weeks, quebracho extracts were administered per os for 8 days to one group. A comparable group which did not receive tannins was included as the control. The kids were slaughtered on week 11. Parasitological and pathophysiological parameters were measured weekly. Worm counts were assessed and mast cells, globule leukocytes and eosinophils were counted in the abomasal and intestinal mucosae. Tannin administration was associated with a decrease in egg excretion, and a decrease in female fecundity, but with no changes in worm numbers. These changes were associated with an increased number of intestinal mast cells. In experiment 2, 24 goats were used according to a 2 x 2 factorial design, depending on infection and tannin administration. Two groups were either infected with 6 000 L3 of T. colubriformis or T. circumcincta. Within each group, the goats were either drenched or undrenched with tannin extracts. Pathophysiological parameters were measured weekly. Twelve days after the cessation of tannin administration, the goats were slaughtered. Worm counts and female worm fecundity were determined. Tannin consumption was associated with a significant reduction (P < 0.01) of Trichostrongylus populations and a close to significant reduction for Teladorsagia. No effect on fecundity was observed. Our results (1) confirm the consequences of condensed tannins on nematodes in goats as in sheep and (2) indicate divergent effects depending on the parasitic stage exposed to the condensed tannins.

Polymorphonuclear proteolytic activity and milk composition change

Abstract: Relationships between Quarter Milk Cell Count (QMCC) and Tank Milk Cell Count (TMCC) with milk biochemical and technological parameters in milk and dairy products were investigated. All parameters measured were affected by the increase of TMCC and QMCC between 0 and 600 000 cells/mL. The variable effect of lactation stage which is different for different authors, is discussed. The three mechanisms, measured during the inflammation of the udder, implicated in the modification of milk quality are described (a decrease in synthesis, a decrease in the milk barrier permeability and an increase in proteolytic activities). The direct effect of plasmin in caseinolysis is well known; the specific role of the increase of somatic cells (especially PMN) in the modification of milk quality is described. Several specific proteolytic activities of PMN are described and the impact of these activities on caseinolysis is evaluated. Two hypothetical mechanisms of caseinolysis by PMN are suggested and a synthetic scheme of the role of plasmin, bacteria and somatic cells in caseinolysis is discussed.

Apoptosis in the lungs of pigs infected with porcine reproductive and respiratory syndrome virus and associations with the production of apoptogenic cytokines.

Abstract: Apoptosis was studied in the lungs of pigs during an infection with a European strain of porcine reproductive and respiratory syndrome virus (PRRSV) and it was examined if cytokines were involved in the induction of apoptosis. Twenty-two 4- to 5-week-old gnotobiotic pigs were inoculated intranasally with 10(6.0) TCID50 of the Lelystad virus and euthanised between 1 and 52 days post inoculation (PI). The lungs and broncho-alveolar lavage (BAL) cells were assessed both for virus replication and apoptosis; BAL fluids were examined for interleukin (IL)-1, tumour necrosis factor-alpha and IL-10. Double-labellings were conducted to determine the relation between virus replication and apoptosis and to identify the apoptotic cells. Apoptosis occurred in both infected and non-infected cells. The percentages of infected cells, which were apoptotic, ranged between 9 and 39% in the lungs and between 13 and 30% in the BAL cells. The majority of apoptotic cells were non-infected. Non-infected apoptotic cells in the lungs were predominantly monocytes/macrophages, whereas those in the broncho-alveolar spaces were predominantly lymphocytes. The peak of apoptosis in the lungs at 14 days PI was preceded by a peak of IL-1 and IL-10 production at 9 days PI, suggesting a possible role of these cytokines in the induction of apoptosis in non-infected interstitial monocytes/macrophages. However, the latter hypothesis was not confirmed in vitro, since blood monocytes or alveolar macrophages did not undergo apoptosis after treatment with recombinant porcine IL-1 or IL-10.

Multispecies and multiple anthelmintic resistance on cattle nematodes in a farm in Argentina: the beginning of high resistance?

Abstract: There are very few resistance records on cattle nematodes. South American successive records have been increasing rapidly since the year 2000. In Argentina, increasing dissatisfaction based on the exclusive use of macrocyclic lactones has prompted the use of benzimidazoles in the Pampean region. The studied farm is located in the Argentina humid Pampas and had apparently poor results after anthelmintic treatments. Evaluation of resistance was firstly based on faecal egg reduction after treatment in November 2001 and June 2002 and was complemented with worm counts obtained by necropsy in June 2002. The study reports that the reduction of faecal egg excretion after benzimidazole (reduction 31-79%) or ivermectin treatments (76-97%) in November 2001 was insufficient and these poor results were confirmed in June 2002 (benzimidazoles (65- 89%) and ivermectin (47-77%)). Several methods for evaluating faecal egg count reduction were performed and yielded different results. A new method based on the estimated faecal egg counts (corrected from the initial faecal egg counts and animal type using a general linear model) was used and gave higher flexibility in the interpretation of putative resistance to anthelmintics. From necropsy results, it could be concluded that Cooperia oncophora was resistant to avermectins and benzimidazoles and that Cooperia punctata, Ostertagia ostertagi and Haemonchus placei were resistant to benzimidazoles. This case of multispecies and multidrug resistance is probably not unique and could reflect the emergence of resistances in Argentina. This emergence is probably due to the intensive use of anthelmintics, the absence of refugia, and the frequent circulation of infected cattle. nematode / cattle / resistance / anthelmintic / Argentina

The complement in milk and defense of the bovine mammary gland against infections.

Abstract: The mammary gland of dairy cows, which is prone to infection by various bacteria, mobilizes local and systemic immune defenses to cope with pathogens. The complement system plays an important part in the innate immunity against microorganisms through its bactericidal, opsonic, and phlogistic functions. The amount of the complement in the milk of healthy glands of dairy cows is low. Moreover, the classical pathway of activation is not functional because of a shortage in C1q. By contrast, the alternative pathway is active, deposits C3b and C3bi on bacteria, and generates amounts of C5a which are highly variable among cows. A slight inhibition of the bactericidal/hemolytic activities, of the deposition of C1q on bacteria, and of the phlogistic activity of C5a makes milk a rather anti-inflammatory fluid. The inhibitory activity does not involve C3b/ C3bi deposition on bacteria, nor the generation of C5a by the alternative pathway. When inflammation develops, the blood-derived complement components overcome the inhibitions and complement-dependent bactericidal, opsonic and phlogistic activities may be high in milk. Further research is necessary to evaluate the contribution of C5a to the recruitment of leukocytes in the mammary gland, and to specify the links between the complement system and the response of resident cells (leukocytes and mammary epithelial cells) to infection stimulus. This will help to define the contribution of the complement system to resistance against mastitis, and could help to differentiate animals more or less resistant to this frequent and costly disease.

Left displacement of the abomasum in dairy cattle: recent developments in epidemiological and etiological aspects

Abstract: The research with respect to displacement of the abomasum (DA) in dairy cattle is reviewed. Evaluated articles describe epidemiological and experimental studies. The occurrence is elevated with regard to breed, gender, age, concurrent diseases, environmental aspects and production levels as contributing factors and emphasis is placed on the effects of nutrition and metabolism. Reviewing the experimental work, distinction is made between the research into gas production in the abomasum and hypomotility of the abomasum, since both represent presumed pathways in the development of DA. Although the different fields of research have positive contributions to the understanding of the pathogenesis of DA, contradictions in the different studies are present. This is partly due to extrapolation of results from sheep to cows, or because of a low number of cows in the experiments. Finally, general suggestions are made for further research in the field of the pathogenesis of DA.

The current status of major tick borne diseases in Zambia

Abstract: Tick-borne diseases occurring in Zambia are assuming more importance as they continue to be a major economic problem not only in Zambia, but in many parts of Eastern, Southern and Central Africa. The current control methods, which include the use of toxic acaricides to kill ticks, and the virulent sporozoite infection and treatment method have limitations. Recombinant vaccines, currently in their experimental stages, offer hope for the future. The use of acaricides is hampered by the development of acaricide resistance and live vaccines are dependent on cold chain facilities, which are a formidable obstacle in the poorly developed infrastructure in parts of Zambia where the vaccine is most needed. Amidst these drawbacks are the results of the recent research on parasites and vector recombinant vaccines which promise to circumvent these problems. The history, current status and attitudes regarding the control of these diseases, taking into account their complexity, are reviewed. The establishment of the well-designed Central Veterinary Research Institute (CVRI) and Japanese International Cooperation Agency (JICA) sponsored veterinary school, both have a potential for high quality research, with access to a wealth of specimens a veritable goldmine of research material. It is thus hoped that this review will stimulate the desire to maximize the value of the tick and tick-borne disease research in both Zambia and the international research community.

Influence of age and purpose for testing on the cut-off selection of serological methods in bovine neosporosis

Abstract: The aim of this study was to investigate the need for different cut-off points, according to animal age and the purpose of testing, for two of the most widely used serological techniques in bovine neosporosis, IFAT and a crude antigen ELISA (Civtest®, HIPRA). Therefore, the population reference sera used were defined using a combination of multiple criteria such as epidemiological/ clinical and histopathological parameters and an immunoblot test. Firstly, foetuses and breeding cattle (heifers and cows) were considered as separate subpopulations for serological evaluation. Secondly, cut-off points for each serological technique (IFAT and ELISA) according to age group (foetuses and breeding cattle) and the different practical applications (detection of infection and abortion) were calculated following the receiver operating characteristics (ROC) analysis. Cut-off points were defined, for IFAT and ELISA for aborted breeding cattle and for IFAT alone in the case of the foetuses, assuming an equivalent cost of false positive and negative results. In infected breeding cattle, for IFAT and ELISA and in foetuses for ELISA, two possible cut-off values were obtained, one for a maximum sensitivity and one for a maximum specificity and the intervals of unclear results were defined. In this case, a cut-off value for equal sensitivity and specificity was also estimated. When cut-off points for infected breeding cattle, 1:100-1:250 for IFAT and 0.306- 0.451 for ELISA were applied to a target population, optimal and similar negative and positive predictive values together with similar apparent and true prevalence results were observed suggesting the possibility of using both tests interchangeably. Neospora caninum / foetus / breeding cattle / serology / cut-off

Classification of Dutch and German avian reoviruses by sequencing the sigma-C protein.

Abstract: We have amplified, cloned and sequenced (part of) the open reading frame of the S1 segment encoding the ? C protein of avian reoviruses isolated from chickens with different disease conditions in Germany and The Netherlands during 1980 up to 2000. These avian reoviruses were analysed phylogenetically and compared with sequences of avian reoviruses in the Genbank database. The avian reoviruses could be grouped in 5 different genotyping clusters and this classification was identical when the sequences were compared of the 5? end, the 3? end or the whole open reading frame of the ? C protein. Therefore sequencing of either part of the gene encoding the ? C protein seems to be reliable for classification. We were unable to identify a correlation between ? C sequences of the avian reoviruses and the disease condition they were isolated from. The sequences found in The Netherlands and in Germany are, like those in Taiwan, more dispersed than the known avian reovirus ? C sequences in the USA and Australia. We did not establish temporal or geographic differences in the avian reoviruses studied.

Evaluation of a modified Rose Bengal test and an indirect enzyme-linked immunosorbent assay for the diagnosis of Brucella melitensis infection in sheep.

Abstract: A modified Rose Bengal test (mRB) and an indirect ELISA (iELISA) with Protein G as the conjugate, were evaluated for the diagnosis of Brucella melitensis infection in unvaccinated sheep with a known bacteriological status, and their diagnostic efficacy was compared with that of the standard Rose Bengal (RB) and Complement Fixation (CF) tests used in the current eradication campaign in EU countries. All tests showed 100% specificity when testing the sera from 212 Brucella-free sheep. When testing the sera from 219 Brucella melitensis culture-positive sheep, both the mRB and iELISA tests were more sensitive (98.6% and 96.8%, respectively) than the RB and CF tests (95.0% and 92.7%, respectively). These results were similar when testing the sera from 181 animals belonging to infected flocks but found bacteriologically negative, suggesting that the mRB or iELISA tests could advantageously replace the current RB procedure used as the screening test.

A pharmacokinetic model to document the actual disposition of topical ivermectin in cattle.

Abstract: Ivermectin is a worldwide-used antiparasitic drug largely administered to cattle as a topical formulation (pour-on). The actual plasma and faecal disposition of pour-on ivermectin in cattle was documented using an original pharmacokinetic model, and taking into account the oral ingestion of the topical drug following physiological licking as a secondary route of exposure. Six pairs of monozygotic twin cattle received successively one i.v. and two pour-on administrations of ivermectin at a 3-5-month interval. For one pour-on administration, the twins were separated into an unrestrained group and a group where self- and allo-licking were prevented. Ivermectin concentrations in the plasma and faeces were determined by HPLC. Licking resulted in a high intra-and inter-individual variability of systemic exposure after topical application. By the means of pharmacokinetic modelling, we showed that 58-87% of the pour-on dose was ingested, while only 10% was absorbed percutaneously. Approximately 72% of the ingested ivermectin transited directly into the faeces, resulting in a 7-fold higher faecal excretion of the parent drug than in the non-lickers. We conclude that topical administration does not guarantee a controlled drug delivery in cattle. More importantly, the simulations revealed that non-treated cattle could get easily contaminated by allo-licking, raising the public health problem of unexpected drug residues in edible tissues.

Milk transfer of cyanide and thiocyanate: Cyanide exposure by lactation in goats.

Abstract: The present work was aimed at evaluating the effects of maternal exposure to potassium cyanide (KCN) during lactation in goats. Twenty-seven lactating female goats were orally dosed with 0 (control), 1.0, 2.0, or 3.0 mg KCN/kg body weight/day from lactation days 0 to 90. After this period, all male kids and one mother from each group were killed for a pathological study. Cyanide treatment promoted the clinical signs of maternal toxicity in the highest KCN group but did not affect body weight. Both cyanide and thiocyanate presented increased levels in both dams and kids from the treated groups. Microscopic lesions, but without alterations on the biochemical panel, were found in the brain, thyroid, liver, and kidneys of both dams and kids from the treated groups. These findings suggest that lactating offspring can be indirectly intoxicated by maternal exposure to cyanide.

Cellular and humoral local immune responses in sheep experimentally infected with Oestrus ovis (Diptera: Oestridae)

Abstract: Cellular and humoral local responses were investigated following repetitive artificial Oestrus ovis infections in lambs The presence of larvae induced a huge local recruitment of either leucocytes (T and B lymphocytes, macrophages) or granulocytes (eosinophils, mast cells and globule leucocytes) This cellular response was more pronounced in the ethmoid and sinus (development sites of second and third instar larvae) than in the septum or turbinates where first instar larvae migrate Infected lambs produced Oestrus ovis specific IgG and IgA antibodies in their mucus This local humoral response was mainly directed against larval salivary gland antigens and not against larval digestive tract antigens Compared to the control animals, the sinusal mucosa of infected animals was extremely thickened and the epithelium exhibited hyperplasia, metaplasia and eosinophilic exocytosis The possible roles of these local immune responses in the regulation of O ovis larvae populations in sheep are discussed

Recombinant bovine soluble CD14 reduces severity of experimental Escherichia coli mastitis in mice.

Abstract: Endotoxin, or lipopolysaccharide (LPS), is responsible for pathogenesis of infections induced by Gram-negative bacteria, such as E. coli. The cellular response to LPS is modulated by interactions among LPS, LPS-binding protein (LBP) and CD14. Accumulated evidence shows that the soluble form of CD14 (sCD14) competes with membrane-bound CD14 (mCD14) for LPS and plays a pivotal role in regulating bacterial infection and septic shock caused by Gram-negative bacteria. Recombinant bovine sCD14 (rbosCD14) was produced by transfected insect sf/9 cells and its biological function was evaluated in mice. Eighty-one 8-week old BALB/cj female mice were randomly assigned to two groups, and injected intraperitoneally with either LPS (8 microg/g of body weight, n = 41) or LPS plus rbosCD14 (6.8 microg/g of body weight, n = 40). Survival rate at 24 h after injection for mice injected with either LPS or LPS plus rbosCD14 was 30 and 72%, respectively (P < 0.01). At 48 h survival rate was 7 and 37%, respectively (P < 0.01). To investigate the protective effect of rbosCD14 on experimentally induced mastitis in mice, two abdominal contralateral mammary glands of 7 lactating BALB/cj mice were injected through the teat canal with 10-20 colony-forming units (CFU) of Escherichia coli. One gland simultaneously received rbosCD14 (6 microg) and the other saline. At 24 h after challenge, glands that received rbosCD14 had less swelling and hemorrhaging, significantly lower bacterial counts (P < 0.05) and lower concentrations of TNF-alpha (P < 0.05). Results indicate that rbosCD14 is biologically functional and reduces mortality in mice from endotoxin shock and severity of intramammary infection by E. coli.

Detection of foot-and-mouth disease virus from culture and clinical samples by reverse transcription-PCR coupled to restriction enzyme and sequence analysis.

Abstract: A reverse transcription-PCR (RT-PCR) method is presented for the highly sensitive and specific detection of foot-and-mouth disease virus (FMDV). A primer pair flanking a region of the viral polymerase gene (3D) corresponding to the C-terminus of the protein was designed and a single step RT-PCR reaction was developed. The assay allowed the detection of viral RNA from a variety of animal samples and from a wide range of FMDV isolates of different origins and serotypes. The presence of an Ahd I restriction site within the amplicon in 96% of the isolates analyzed allowed an additional confirmation step of the positive reactions by a simple digestion yielding characteristic fragment sizes. The set of primers described here was suitable for direct sequencing of the PCR product (290 bp), and the nucleotide sequences corresponding to the SAT 1 and SAT 3 strains were determined. The segment amplified, when used in phylogenetic studies, allowed the clustering of SAT isolates and the rest of FMDV strains as two separate lineages.

Comparison of genome segments 2, 7 and 10 of bluetongue viruses serotype 2 for differentiation between field isolates and the vaccine strain

Abstract: Bluetongue (BT) virus serotype 2 (BTV 2) was first confirmed in Tunisia in February 2000 and has since spread northward and westward, infecting several other countries and islands, including Corsica, where clinical disease was reported in October 2000. BT was again reported on the Island in July 2001, some six months after a vaccination campaign against BTV 2. The molecular relationship between isolates of the BTV 2 Corsican wild-type viruses from 2000 and 2001, and the attenuated BTV 2 vaccine were determined by comparing corresponding sequences of genome segments 2, 7 and 10 with each other and with already published sequences available in the genome database. Complete genetic stability was observed between the isolates of the Corsican BTV 2. There was some divergence between the nucleotide sequences of segment 10 obtained from the wild-type and vaccine virus strains. Based on these differences, primers were selected that could be used in RT-PCR to differentiate between the wild-type and the vaccine viruses.

Humoral and cell-mediated immune responses of d/d histocompatible pigs against classical swine fever (CSF) virus

Abstract: A better understanding of cell-mediated immune responses to classical swine fever virus (CSFV) is essential for the future development of improved vaccines. We analyzed the generation of cell-mediated and humoral immune responses in d/d histocompatible pigs following CSFV infection or vaccination. Viral infection induced high T cell responses with high primary and secondary CTL activity correlated with high IFN-gamma production, whereas vaccination with a live vaccine followed by infection mainly induced neutralizing antibody but low cell-mediated responses. Moreover, high IgG1 response was associated with high IFN-gamma response following infection whereas a weak IFN-gamma response was related to a good IgG2 response but a low IgG1 production. These data could reflect Th1/Th2-like balance of immune responses depending upon immunization protocols, which has not yet been described in the pig. T-cell responses to CSFV were evidenced by CSFV-specific CD25 upregulation on CD4-CD8+, but not on CD4+CD8- cells, which further illustrated the importance of CTL responses after infection. Our results indicated that generation of cell-mediated immune responses was much higher following intranasal/oral CSFV infection than after intramuscular vaccination, which implies that the capacity of new CSFV vaccines to induce higher T-cell responses should be considered.

SDS-PAGE and Western blot of urinary proteins in dogs with leishmaniasis.

Abstract: Canine leishmaniasis is an endemic disease in the Mediterranean area caused by the protozoan Leishmania infantum, which usually produces renal failure. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and Western blot using antibodies to IgG and IgA from dogs were carried out in the urine of 22 dogs with leishmaniasis diagnosed by ELISA and confirmed by PCR, and 20 healthy dogs. The results were compared to renal function laboratory tests and to those from a histopathological study of the kidneys from sick animals that died naturally or were euthanized. Five different bands with molecular weights ranging from 10 to 110 kDa were obtained from the electrophoresis of the urine of healthy dogs. 33.5% of total proteins corresponded to low molecular weight proteins and the other proteins had middle and high molecular weights. However, in the group with leishmaniasis, a maximum of 11 different bands with molecular weights ranging from 10 kDa to 150 kDa were displayed in the electrophoresis of the urine. The urine electrophoretic pattern in the sick dogs was classified as mixed (proteins with high and low molecular weights) because low molecular weight proteins made up 57.9% and the rest of the proteins had middle and high molecular weights. In Western blot, none of the healthy dogs showed excretion of IgG and/or IgA, whereas IgG and IgA were detected in the Western blot of urine of 68% and 55% respectively of dogs with leishmaniasis. The results obtained in the leishmaniasis group agreed with glomerular and tubular damage, which were confirmed by the histopathological findings.

Expression of potential lymphocyte trafficking mediator molecules in the mammary gland

Abstract: The mammary gland performs a variety of immunological functions, including protecting itself from mastitis and protecting neonates from infectious agents. Several molecules that mediate lymphocyte trafficking in the immune system are also expressed in the mammary gland. This review is focused on the immunological function of these molecules, especially glycosylation-dependent cell adhesion molecule-1 (GlyCAM-1) and mucosal addressin cell adhesion molecule-1 (MAdCAM-1) in the mammary gland. GlyCAM-1 is expressed in the lactating mouse mammary gland. Endothelial cells produce this protein and secrete it into milk. The glycosylated modification of mammary gland GlyCAM-1 is different from that of the lymph nodes, and lacks the binding ability for L-selectin on lymphocytes. GlyCAM-1 in the mammary gland is not involved in lymphocyte migration, and probably has another function besides that of the lymph nodes. MAdCAM-1 is expressed on endothelial cells of small venules around mouse mammary lobules during lactation. This molecule has the ability to interact with α4s7 integrin on lymphocytes and mediates lymphocyte recruitment to the mammary gland. The density of s7 + /CD3 + T-cells is correlated with the density of the MAdCAM-1-stained area, suggesting that MAdCAM-1 may mediate the migration of these cells. In contrast, there is no relationship between MAdCAM-1 expression and the number of s 7 + /c-IgA + B-cells, implying that some other factor is involved in lymphocyte migration to the mammary gland. Chemokines, such as IL-8, GRO-α, MCP-1, RANTES and MEC, have been detected in human and mouse mammary glands. Although little information is available, these molecules may contribute to lymphocyte migration to the mammary gland. mammary gland / mucosal-associated lymphoid tissue (MALT) / glycosylation-dependent cell adhesion molecule-1 (GlyCAM-1) / mucosal addressin cell adhesion molecule-1 (MAdCAM-1) / lymphocyte migration

Listeria monocytogenes contamination of finishing pigs: an exploratory epidemiological survey in France

Abstract: Listeria monocytogenes is a foodborne pathogen of major concern for public health in industrialised countries. Since L. monocytogenes carriage by pigs at the herd level could be a primary source for carcass contamination, control measures should be designed to reduce the L. monocytogenes load at the pre-harvest stage. For this purpose, an exploratory analytical survey was carried out in 2000-2001 in 93 French farrow-to-finish pig farms concerning L. monocytogenes contamination in pigs before they left for the slaughterhouse. On each farm, the L. monocytogenes status of a batch of contemporary fattening pigs housed in the same room was assessed on faecal material samples taken by means of gauze swabs wiped on the perianal region of the pigs. Fourteen percent of the batches studied had at least one contaminated sample and were therefore classified as L. monocytogenes contaminated batches. Two logistic regression models were used to assess the association between managerial and hygiene practices and the risk of L. monocytogenes contamination of the batch at the end of the finishing period on the whole data set (n = 93) and in the wet feeding farms only (n = 57). Wet feeding during the fattening period was identified as a risk factor for L. monocytogenes contamination. Risk factors related to the introduction of L. monocytogenes in pig facilities were identified for both the general and wet feeding farm data sets. Poor care paid to hygiene on the farms was found to increase the risk of being infected (boots cleaning, change room presence). When the duration of the empty period prior to the introduction of growing pigs was less than one day in the fattening section, the risk of L. monocytogenes contamination was significantly increased. For wet feeding farms, a distribution pipeline cleaning procedure including disinfection was found to be associated with a higher risk of contamination than no cleaning or a procedure consisting of rinsing with water only.