Showing papers in "Journal of Veterinary Medical Science in 2012"
TL;DR: In this paper, four methods were evaluated for exosome isolation from bovine milk: ExoQuick precipitation, ultracentrifugation with exoquick precipitation and density gradient centrifugation.
Abstract: Four methods were evaluated for isolating exosomes from bovine milk: (1) ExoQuick precipitation, (2) ultracentrifugation with ExoQuick precipitation, (3) ultracentrifugation with density gradient centrifugation, and (4) human milk exosome isolation. Methods 1 and 4 failed due to differences between bovine and human milk. Exosomes were efficiently isolated by ultracentrifugation with either ExoQuick precipitation (method 2) or density gradient centrifugation (method 3). The highest yield of exosomes was achieved using ultracentrifugation with ExoQuick precipitation, whereas higher quality exosome isolation with intact morphological structures was achieved by ultracentrifugation with density gradient centrifugation.
133 citations
TL;DR: The data suggest that transplantation of MSCs promotes functional recovery after SCI, and application of UCSCs led to more nerve regeneration, neuroprotection and less inflammation compared to other M SCs.
Abstract: Previous animal studies have shown that transplantation of mesenchymal stem cells (MSCs) into spinal cord lesions enhances axonal regeneration and promotes functional recovery. We isolated the MSCs derived from fat, bone marrow, Wharton's jelly and umbilical cord blood (UCB) positive for MSC markers and negative for hematopoietic cell markers. Their effects on the regeneration of injured canine spinal cords were compared. Spinal cord injury was induced by balloon catheter compression. Dogs with injured spinal cords were treated with only matrigel or matrigel mixed with each type of MSCs. Olby and modified Tarlov scores, immunohistochemistry, ELISA and Western blot analysis were used to evaluate the therapeutic effects. The different MSC groups showed significant improvements in locomotion at 8 weeks after transplantation (P<0.05). This recovery was accompanied by increased numbers of surviving neuron and neurofilament-positive fibers in the lesion site. Compared to the control, the lesion sizes were smaller, and fewer microglia and reactive astrocytes were found in the spinal cord epicenter of all MSC groups. Although there were no significant differences in functional recovery among the MSCs groups, UCB-derived MSCs (UCSCs) induced more nerve regeneration and anti-inflammation activity (P<0.05). Transplanted MSCs survived for 8 weeks and reduced IL-6 and COX-2 levels, which may have promoted neuronal regeneration in the spinal cord. Our data suggest that transplantation of MSCs promotes functional recovery after SCI. Furthermore, application of UCSCs led to more nerve regeneration, neuroprotection and less inflammation compared to other MSCs.
115 citations
TL;DR: The Foot-and-mouth disease epidemic in Japan was contained within a localized area in Miyazaki Prefecture and was eradicated within three months because of intensive control efforts including emergency vaccination.
Abstract: Foot-and-mouth disease (FMD) occurred recently for the first time in a decade in Japan. The index case was detected on a beef-breeding farm in Miyazaki Prefecture, Southern Japan, on April 20, 2010. After confirmation of this first case, control measures such as stamping out, movement restriction and disinfection were implemented. However, these strategies proved insufficient to prevent the spread of FMD and emergency vaccination was adopted. Up until the last outbreak on July 4, 2010, a total of 292 outbreaks had been confirmed, with about 290,000 animals having been culled. The epidemic occurred in an area with a high density of cattle and pigs, making disease control difficult. Invasion of the disease into a high-density area aided its rapid spread and led to difficulties in locating suitable burial sites. Epidemiological investigations indicated that the disease was introduced into Japan approximately one month before detection. This delay in initial detection is considered to have allowed an increased number of outbreaks in the early stage of the epidemic. Nevertheless, the epidemic was contained within a localized area in Miyazaki Prefecture and was eradicated within three months because of intensive control efforts including emergency vaccination. Although this epidemic devastated the livestock industry in Japan, many lessons can be learnt for the future prevention and control of infectious diseases in animals.
108 citations
TL;DR: Evidence is provided that the proliferation of CTX-M-producing E. coli is due to the growth of indigenous CTX,M- producing strains and the possible emergence of strains that acquired CTx-M genes by horizontal transfer in different broiler farms, which should be controlled.
Abstract: To evaluate the diversity of extended-spectrum β-lactamases (ESBL) genes among food-producing animals, 48 isolates of ESBL-producing Escherichia coli isolates were obtained from rectal samples of broilers, layers, beef cattle and pigs, at the slaughterhouse level. ESBL-carrying E. coli were isolated from 60.0% of individual broiler rectal samples, 5.9% of layers, 12.5% of beef cattle and 3% of pigs. One ESBL-producing Klebsiella pneumoniae was isolated from a broiler. The ESBL-positive E. coli isolates from broilers harbored various ESBL genes: bla (SHV-12), bla(CTX-M-2), bla(CTX-M-14), bla(CTX-M-15) and bla(CTX-M-44). The plasmid DNAs were analyzed by restriction patterns. Homogeneous band patterns were yielded in those of K. pneumoniae and E. coli isolates harboring the bla(CTX-M-2) gene from different farms. No genetic relation between the 2 CTX-M-14 ESBL-producing strains was found by pulsed-field gel electrophoresis, although 2 plasmids in these strains, obtained from different broiler farms, were similar to each other. This study provides evidence that the proliferation of CTX-M-producing E. coli is due to the growth of indigenous CTX-M-producing strains and the possible emergence of strains that acquired CTX-M genes by horizontal transfer in different broiler farms. CTX-M-producing coliforms in broilers should be controlled due to the critical importance of cephalosporins and the zoonotic potential of ESBL-producing bacteria.
100 citations
TL;DR: In most cases, VBNC cells failed to resuscitate in Caco-2 cells, but in some experiments, they formed colonies after co-incubation with host cells, and C. jejuni remains virulent which may possibly lead to disease in humans.
Abstract: Campylobacter jejuni is the major cause of human gastroenteritis worldwide. Under stress conditions, C. jejuni can enter a viable but non-culturable (VBNC) state. We found that the C. jejuni was able to enter a VBNC state by prolonged incubation at 4°C. The standard isolation methods using pre-enrichment steps in Bolton broth or Preston broth could not detect the VBNC cells in spiked chicken meat. The transcription levels of virulence-associated genes (flaA, flaB, cadF, ciaB, cdtA, cdtB and cdtC) were expressed in VBNC cells but in low levels. The VBNC cells retained the ability to invade Caco-2 human intestinal epithelial cells in vitro. In most cases, VBNC cells failed to resuscitate in Caco-2 cells, but in some experiments, they formed colonies after co-incubation with host cells. Collectively, C. jejuni enters into a VBNC state at 4°C and the VBNC C. jejuni remains virulent which may possibly lead to disease in humans. C. jejuni in VBNC state is a potential concern for food safety.
82 citations
TL;DR: The findings indicate that curcumin specifically binds to the aggregated Aβ molecules in various animals, and further to phosphorylated tau protein, probably according to its conformational nature.
Abstract: The binding of curcumin to senile plaques (SPs) and cerebral amyloid angiopathy (CAA) was examined in the aged brain of various animal species and a human patient with Alzheimer's disease (AD), together with its binding to neurofibrillary tangles (NFTs). Brain sections were immunostained with anti-amyloid β protein 1-42 (Aβ42) and anti-amyloid β protein 1-40 (Aβ40) antibodies. These sections were also stained with alkaline Congo red, periodic acid-methenamine silver (PAM), and curcumin (0.009% curcumin solution) with or without formic acid pretreatment. The sections from the AD brain were also immunostained for anti-paired helical filament-tau (PHF-tau), and were stained with Gallyas silver for NFTs. Some SPs in the AD, monkey, dog, bear, and amyloid precursor protein transgenic mouse (APP Tg-mouse) brains contained congophilic materials, and were intensely positive for curcumin. In addition, curcumin labeled some diffuse SPs negative for Congo red in the AD, monkey, bear, and APP Tg-mouse brains. In all animals, CAA was intensely positive for both Congo red and curcumin. The specific curcumin staining activity was lost by formic acid pretreatment. In the AD brain, NFTs positive for PHF-tau and Gallyas silver were moderately stained with curcumin. These findings indicate that curcumin specifically binds to the aggregated Aβ molecules in various animals, and further to phosphorylated tau protein, probably according to its conformational nature.
67 citations
TL;DR: Phylogenetic analysis based on the part of the hexon gene that included the L1 region revealed that all isolates were almost identical except one isolate in 2009, which suggests a common ancestor for the FAdVs obtained from chickens with IBH in Japan in 2010.
Abstract: Nine fowl adenoviruses (FAdVs) isolated from chickens with inclusion body hepatitis (IBH) in Japan from 2009 to 2010 were characterized serologically and genetically. These isolates were all neutralized by antisera against the SR-48 strain (FAdV-2). Phylogenetic analysis based on the part of the hexon gene that included the L1 region revealed that all isolates were almost identical except one isolate in 2009. This suggests a common ancestor for the FAdVs obtained from chickens with IBH in Japan in 2010.
56 citations
TL;DR: Findings suggest that miR-145 acts as a tumor suppressor in both canine and human malignant melanomas.
Abstract: MicroRNA-145 (miRNA-145; miR-145) is aberrantly expressed in most of human cancers and plays a significant role in carcinogenesis and cancer progression. In the current study, we focused on how miR-145 plays a role in canine and human malignant melanomas. MiR-145 was significantly downregulated in canine malignant melanoma tissues and canine melanoma cell lines, as well as human melanoma cell lines tested. The ectopic expression of miR-145 showed a significant growth inhibition in both canine and human melanoma cells tested, and the effect was achieved partly by suppressing c-MYC in canine melanoma LMeC and in human melanoma A2058 and Mewo cells. At the same time, a suppressive tendency on cell migration in canine melanoma KMeC cells and significant suppression of cell migration in human melanoma A2058 cells by suppressing FASCIN1 were also found. These findings suggest that miR-145 acts as a tumor suppressor in both canine and human malignant melanomas.
56 citations
TL;DR: Seropositivity of lions in the zoo and wild bears captured around this area supported the theory that a CDV epidemic had occurred in many mammal species in and around the zoo, and results indicate a risk of CDV transmission among many animal species, including large felids and endangered species.
Abstract: Canine distemper virus (CDV), a morbillivirus that causes one of the most contagious and lethal viral diseases known in canids, has an expanding host range, including wild animals. Since December 2009, several dead or dying wild raccoon dogs (Nyctereutes procyonoides) were found in and around one safari-style zoo in Japan, and CDV was isolated from four of these animals. In the subsequent months (January to February 2010), 12 tigers (Panthera tigris) in the zoo developed respiratory and gastrointestinal diseases, and CDV RNA was detected in fecal samples of the examined tigers. In March 2010, one of the tigers developed a neurological disorder and died; CDV was isolated from the lung of this animal. Sequence analysis of the complete hemagglutinin (H) gene and the signal peptide region of the fusion (F) gene showed high homology among these isolates (99.8-100%), indicating that CDV might have been transmitted from raccoon dog to tiger. In addition, these isolates belonged to genotype Asia-1 and had lower homology (<90%) to the vaccine strain (Onderstepoort). Seropositivity of lions (Panthera leo) in the zoo and wild bears (Ursus thibetanus) captured around this area supported the theory that a CDV epidemic had occurred in many mammal species in and around the zoo. These results indicate a risk of CDV transmission among many animal species, including large felids and endangered species.
48 citations
TL;DR: The results of this study indicate that miniature dachshunds are predisposed to develop inflammatory colorectal multiple polyps, for which immunosuppressive therapy may be a treatment option.
Abstract: Medical records of dogs with colorectal polyps were retrospectively reviewed, and clinical presentation of inflammatory colorectal polyps in miniature dachshunds was evaluated. Of 33 dogs found to have colorectal polyps, miniature dachshunds were markedly over-represented with 16 dogs (48%), of which 12 (75%) were found to have inflammatory polyps. Multiple polyps localized between the rectum and the descending colon was the most common finding in miniature dachshunds with inflammatory polyps. Twenty dogs (80%) out of 25 miniature dachshunds with inflammatory colorectal polyps responded to immunosuppressive therapy using prednisolone and cyclosporine. The results of this study indicate that miniature dachshunds are predisposed to develop inflammatory colorectal multiple polyps, for which immunosuppressive therapy may be a treatment option.
43 citations
TL;DR: Findings suggest that the rOmp28 of B. abortus might be a good candidate for developing serological diagnostic tools for bovine brucellosis.
Abstract: Brucella spp. are Gram-negative, facultative, intracellular coccobacilli that are pathogenic to a variety of mammals, including ruminants and humans. The conventional serological test for diagnosing brucellosis in cattle in Korea is the standard tube agglutination test. However, agglutination tests sometimes give false-positive results due to cross-reactions with other pathogens. The outer membrane proteins of Brucella species have been extensively studied for their immunogenicity and serodiagnostic applications. However, an application of B. abortus OMPs for serodiagnosis has not been successfully established. In this study, cloning and expression of B. abortus Omp28, a group 3 antigen, were accomplished by PCR amplification cloning into a pMAL expression system, and purification of a recombinant Omp28 (rOmp28). The immunogenicity of rOmp28 was confirmed by Western blot analysis with Brucella-positive bovine serum. To determine whether rOmp2 has a potential benefit for use in the serodiagnosis of bovine brucellosis, rOmp28-based ELISA and latex bead agglutination test were performed. B. abortus positive (n=122) or negative (n=88) from TAT were positive (118/122, 96.7%) or negative (84/88, 95.4%) in ELISA and were positive (94/122, 77%) or negative (71/88, 81.7%) in that the latex bead agglutination test, respectively. The sensitivity, specificity and accuracy were 96.7, 95.4, 96.2% in ELISA and 77, 80.6, 78.5% in latex bead agglutination test, respectively. These findings suggest that the rOmp28 of B. abortus might be a good candidate for developing serological diagnostic tools for bovine brucellosis.
TL;DR: This work examined whether infection of LCMV-GP-pseudotypes was affected by the expression of four cell surface molecules-Axl and Tyro3 and DC-SIGN and LSECtin-that are known receptors of Lassa virus, another arenavirus.
Abstract: The glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV), the prototype arenavirus, is a promising envelope protein of lentiviral pseudotype vectors for gene therapy. The distribution of dystroglycan, a known receptor for LCMV, cannot explain the narrow tropism of LCMV-GP-pseudotypes. Here, we examined whether infection of LCMV-GP-pseudotypes was affected by the expression of four cell surface molecules-Axl and Tyro3 (from the TAM family) and DC-SIGN and LSECtin (from the C-type lectin family)-that are known receptors of Lassa virus, another arenavirus. All four molecules enhanced LCMV-GP-pseudotype infection of cells. These results help explain the tropism of LCMV-GP-pseudotypes and further our understanding of LCMV infection in animals.
TL;DR: Gentamycin, streptomycin and tetracycline may be suitable antibiotics for the treatment or control of disease caused by Arcobacter spp.
Abstract: The objective of this study was to determine the antibiotic susceptibility of Arcobacter spp. isolated from various sources. Seventy Arcobacter spp. isolates were tested for their susceptibility to 13 antimicrobial agents. Antimicrobial susceptibility testing was performed by using the agar disc diffusion method on Mueller-Hinton agar supplemented with 5% defibrinated sheep blood. The antibiotics tested included enrofloxacin, erythromycin, streptomycin, gentamycin, rifampin, tetracycline, ampicillin, trimethoprim/sulfamethoxazole, nalidixic acid, danofloxacin, amoxycillin-clavulonic acid, cefuroxime-sodium, neomycine. Although all the arcobacters tested were susceptible to gentamycin, resistance to three or more antibacterial agents (especially, trimethoprim/sulfamethoxazole, cefuroxime-sodium and rifampin) was observed. A. butzleri isolates were found to be resistant to amoxycillin+clavulonic acid, nalidixic acid and ampicillin, at the rate of 20%, 44.28% and 78.57% respectively. In conclusion, gentamycin, streptomycin and tetracycline may be suitable antibiotics for the treatment or control of disease caused by Arcobacter spp. in veterinary and human medicine.
TL;DR: In this article, the effect of the lytic phage CJ12 in ETEC-infected pigs was evaluated with a 1:1,000 (0.1%) ratio.
Abstract: We evaluated effect of enterotoxigenic Escherichia coli (ETEC) specific lytic phage CJ12 in ETEC infected pigs. Phage was mixed with feed at a ratio of 1:1,000 (0.1%). One week after initially providing phage mixed feed, pigs were challenged orally with 10(11) CFU of ETEC and body weight, diarrhea score, bacterial CFU and phage PFU in the feces were measured. Pigs of phage treated groups C (10(6) PFU/g) and D (10(8) PFU/g) showed more resistance to diarrhea due to ETEC infection compared to positive control group B on the third day after the initial challenge. Moreover, during the quantitation of ETEC in feces, both groups C and D showed approximately 63.92 and 60.73% reduced ETEC compared to positive control group B. Phages were successfully isolated from feces in both groups C and D during the experiment without any adverse effects, suggesting the possibility of using CJ12 as a feed additive.
TL;DR: The possibility that the mongoose is a reservoir animal for HEV on Okinawa Island is emphasized, as 6 HEV RNAs belonged to genotype 3 and were classified into groups A and B.
Abstract: Hepatitis E virus (HEV) infection has previously been reported in wild mongooses on Okinawa Island; to date however, only one HEV RNA sequence has been identified in a mongoose. Hence, this study was performed to detect HEV RNA in 209 wild mongooses on Okinawa Island. Six (2.9%) samples tested positive for HEV RNA. Phylogenetic analysis revealed that 6 HEV RNAs belonged to genotype 3 and were classified into groups A and B. In group B, mongoose-derived HEV sequences were very similar to mongoose HEV previously detected on Okinawa Island, as well as to those of a pig. This investigation emphasized the possibility that the mongoose is a reservoir animal for HEV on Okinawa Island.
TL;DR: It is demonstrated that PHT using indocyanine green (ICG) and near infrared (NIR) light source induces apoptosis and G0/G1 cell cycle arrest in the B16F10 cells.
Abstract: We examined the effects of photodynamic hyperthemal therapy (PHT), which is a combination of photodynamic therapy (PDT) and hyperthermia (HT), on the apoptosis and cell cycle progression of murine melanoma B16F10 cells. The percentage of apoptotic cell was determined by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated Annexin V and propidium iodide (PI) double staining. The cell cycle analysis was performed by PI staining with flow cytometry. The expression of cyclins and heat shock protein 70 (Hsp70) were examined by a Western blotting analysis. PHT induces death in B16F10 cells, and PHT-mediated apoptosis occurred acutely and persistently in vitro. Our study demonstrated that PHT using indocyanine green (ICG) and near infrared (NIR) light source induces apoptosis and G0/G1 cell cycle arrest in the B16F10 cells.
TL;DR: Once daily administration of robenacoxib tablets had noninferior efficacy and tolerability compared to carprofen for the treatment of the clinical signs of osteoarthritis in dogs.
Abstract: The efficacy and tolerability of robenacoxib for the treatment of osteoarthritis in dogs were evaluated in a prospective, multicenter, randomized, noninferiority design clinical trial. A total of 32 dogs presenting with osteoarthritis were allocated randomly to receive, orally once daily for 28 days, either 1-2 mg/kg robenacoxib (n=21) or 3.5-5 mg/kg carprofen (n=11). Dogs were assessed by clinicians and owners using numerical rating scale scores at baseline and days 14 and 28. The primary efficacy endpoint was the global functional disability score, which was the sum of clinician scores for standing posture, lameness at walk, lameness at trot, willingness to raise the contralateral limb and pain at palpation. There was a good to excellent level of efficacy in both treatment groups. Differences between days 14 and 28 compared to day 0 were significant for all 11 clinician and owner scores for robenacoxib, and for 6 of 11 scores for carprofen. The efficacy of robenacoxib was numerically superior to carprofen for all 13 endpoints, but differences were not statistically significant. For the global functional disability score, the estimated efficacy of robenacoxib was 1.244 (95% confidence interval 0.555-2.493) relative to carprofen. The tolerability of both treatments was good as assessed from adverse events, clinical signs, and hematology and serum biochemistry variables. In conclusion, once daily administration of robenacoxib tablets had noninferior efficacy and tolerability compared to carprofen for the treatment of the clinical signs of osteoarthritis in dogs.
TL;DR: Results indicate that PCR and PCR-RFLP using the hypervariable region of HMTp210 are alternative methods to identify the serovar of A. paragallinarum.
Abstract: Avibacterium (Haemophilus) paragallinarum (A. paragallinarum) is a causative agent of infectious coryza in chickens and is classified into three serovars by agglutination tests. In an effort to identify the serovars easily, PCR and PCR-RFLP were employed. As the target gene for PCR, the hypervariable region of HMTp210, which encodes the HA antigen, was used. PCR using primer sets around the hypervariable region amplified 0.8, 1.1 and 1.6 kbp fragments for serovars A, B and C, respectively. Alternatively, the 1.6 kbp fragments were amplified with another primer pair encompassing the hypervariable region and was subjected to digestion with Bgl II, which resulted in the detection of serovar-specific digestion patterns. These results indicate that PCR and PCR-RFLP using the hypervariable region of HMTp210 are alternative methods to identify the serovar of A. paragallinarum.
TL;DR: The data revealed that multidrug resistant Salmonella strains were widely distributed in north Vietnam via the food chain and might contain multiple genes specifying identical resistant phenotypes, thus, continuous studies are necessary to clarify the mechanisms of MDR inSalmonella and its spread in the livestock market.
Abstract: Approximately 39.9% (63/158) of beef samples collected from retail markets in Hanoi from January to June 2009 were Salmonella-positive. Nine Salmonella serovars, Anatum (28.6%), Rissen (25.4%), Weltevreden (12.7%), Typhimurium (7.9%), Derby (7.9%), Lexington (7.9%), Dublin (4.6%), Newport (3.2%) and London (1.8%), were identified. Thirty-seven (58.7%) of the 63 Salmonella isolates were resistant to at least one antimicrobial tested, of which 29 (46%) isolates showed multidrug resistance (MDR). The isolates were commonly resistant to tetracycline (46.0%), sulphonamide (39.7%), ampicilline (31.7%), streptomycin (30.2%), trimethoprim (28.6%), kanamycin (28.6%) and chloramphenicol (22.2%). Fourteen (bla(TEMV), bla(OXA-1), aadA1, aadA2, sul1, tetA, tetB, tetG, cmlA1, floR, dfrA1, dfrA12, aac (3)-IV and aphA1-1AB) out of 22 antimicrobial resistance genes were detected by PCR from the resistant isolates. The catA1, Kn, blaPSE-1 genes and plasmid-mediated quinolones resistance (PMQR) genes such as qnrA, qnrB, qnrS, qepA and acc (6')-ib-cr were not detected. Mutations in the gyrA gene leading to the amino acid changes Ser83Phe and/or Asp87Asn were found in 6 out of the 11 quinolone-resistant isolates. The data revealed that multidrug resistant Salmonella strains were widely distributed in north Vietnam via the food chain and might contain multiple genes specifying identical resistant phenotypes. Thus, continuous studies are necessary to clarify the mechanisms of MDR in Salmonella and its spread in the livestock market.
TL;DR: The results suggest that the occurrence of ESBL-producing E. coli may not be related to feeding with antibiotics and that the contamination of broiler houses with ESBLs might be an important factor.
Abstract: To clarify the factors for occurrence of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli in broilers, two flocks (1 day of age) fed a diet with or without antibiotics were kept in a broiler house sanitized with disinfectants. ESBL-producing E. coli, however, was detected at a concentration of over 10(6) CFU/g of feces at 9 days of age to 49 days of age in both broiler flocks. Therefore, this indicated that the antibiotics other than cephalosporins used in this study had no effect due to co-selection on the numbers of ESBL-producing E. coli in broiler feces during this period. When a flock was kept with diet containing antibiotics for 49 days in a laboratory animal room, no ESBL-producing E. coli was detected in the flock. These results suggest that the occurrence of ESBL-producing E. coli may not be related to feeding with antibiotics and that the contamination of broiler houses with ESBL-producing E. coli might be an important factor.
TL;DR: It is suggested that the use of Matrigel can increase the osteogenic potential of canine Ad-MSCs, a promising source of cells for bone tissue engineering.
Abstract: Adipose tissue-derived mesenchymal stem cells (Ad-MSCs) are a promising source of cells for bone tissue engineering. Matrigel is a basement membrane extract containing multiple extracellular components. This mixture may promote the osteogenic differentiation of MSCs and provide a more appropriate microenvironment for transplanted cells. Here, we investigated the effect of Matrigel on the osteogenic potential of Ad-MSCs. Canine Ad-MSCs were cultured in 2D and 3D matrices and implanted into subcutaneous pouches of dogs either with or without Matrigel. Culture mineralization, cell adhesion efficiency, cell proliferation, osteoid matrix production and alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase activities were quantified and compared. Ad-MSCs grown in 2D cultures with Matrigel showed higher levels of calcium deposition and ALP activity than those grown in the absence of Matrigel under osteogenic conditions. In 3D cultures, the cells cultivated with Matrigel showed greater attachment, proliferation and osteogenic differentiation than those grown without Matrigel. In vivo, Ad-MSCs implanted with Matrigel showed higher osteogenic potential than those without Matrigel. In conclusion, these data suggest that the use of Matrigel can increase the osteogenic potential of canine Ad-MSCs.
TL;DR: The results of the present study suggest that the serotype 1a strains belonging to the group 2 might be widespread in pig populations in Japan.
Abstract: The objective of the present study was to characterize Erysipelothrix sp. strains from recent erysipelas outbreaks in Japan. Eighty-three (100%) strains were identified as E. rhusiopathiae, based on serotyping and spaA PCR. Fifty (60.3%), 5 (6.0%), and 28 (33.7%) strains were isolated from animals with acute, subacute and chronic outbreaks, respectively, of which 79 (95.2%), 1 (1.2%), and 3 (3.6%) belonged to serotypes 1a, 2a, and untypeable, respectively. Fifteen strains (including 3, 2, and 10 from acute, subacute, and chronic cases, respectively) were sensitive to acriflavine, and showed high levels of virulence in mice; of which strains from acute cases, and from subacute and chronic cases killed 100%, and 80 to 100% mice, respectively at challenge doses of 10(2) CFU per mouse. Based on sequence analysis of a 432-bp hypervariable region in spaA gene, 83 strains could be divided into 3 groups: (i) group 1 (3 strains of serotype 1a) had Ala-195 and Ile-203; (ii) group 2 (76 strains of serotype 1a and 3 of untypeable) had Asp-195 and Met-203; and (iii) group 3 (one strain of serotype 2a) had Asn-195 and Ile-203. The results of the present study suggest that the serotype 1a strains belonging to the group 2 might be widespread in pig populations in Japan.
TL;DR: A. hydrophila from farm-raised Nile Telapia serve as a reservoir for antimicrobial resistance determinants as mentioned in this paper, which can be used to detect the presence of class 1, 2 and 3 integrons and all the strains resistant to enrofloxacin and/or ciprofloxinacin.
Abstract: The aim of this study was to determine antimicrobial resistance of Aeromonas hydrophila isolated from farmed Nile Tilapia. A total of 50 A. hydrophila isolates from clinical cases were screened for the presence of class 1, 2 and 3 integrons and all the strains resistant to enrofloxacin and/or ciprofloxacin (n=19) examined for mutation in the quinolone resistance-determining regions (QRDRs) of gyrA and parC. The intI1 gene was detected in 23 A. hydrophila strains (46%) but no intl2 and intl3 were detected. Among these, 14 isolates (60.8%) carried gene cassettes inserted in variable regions i.e., partial aadA2, aadA2, dfrA1-orfC and dfrA12-aadA2, of which the most common gene cassette array was dfrA12-aadA2 (26.09%). Conjugal transfer of class 1 integrons with resistance gene array was detected. All the A. hydrophila strains resistant to enrofloxacin and/or ciprofloxacin possessed mutations in the QRDRs of gyrA and parC. Only a Ser-83-Ile substitution was identified in GyrA and only a Ser-80-Ile amino change was found in ParC. The data confirms that A. hydrophila from farm-raised Nile Telapia serve as a reservoir for antimicrobial resistance determinants.
TL;DR: It was found that the lactose-based extender resulted in a higher percentage of post-thawed sperm motility, viability, intact acrosome and functional plasma membrane than sorbitol-basedextender (P<0.05).
Abstract: The present study determined the effect of different types of sugars (lactose, fructose, glucose and sorbitol) used in egg yolk-based extender on the post-thawed boar semen quality. Twenty-two ejaculates from 6 fertility-proven Yorkshire boars were cryopreserved by liquid nitrogen vapor method. Sperm motility, viability, acrosome integrity and intact functional plasma membrane were determined at 0, 2 and 4 hr after thawing. It was found that the lactose-based extender resulted in a higher percentage of post-thawed sperm motility, viability, intact acrosome and functional plasma membrane than sorbitol-based extender (P<0.05) and fructose-based extender yielded a higher post-thawed sperm motility and viability than sorbitol-based extender (P<0.05). It could be concluded that sorbitol was not an effective sugar for the cryopreservation in boar semen.
TL;DR: Recombinant chicken IFN-λ inhibited influenza virus replication and induced the mRNA expression of the ISGs, Mx and OAS, in chicken embryonic fibroblasts (CEFs) in a dose-dependent manner and showed the potential to induce ISGs and inhibit viral replication in chicken cells.
Abstract: Interferons (IFNs) are key mediators that activate host defense mechanisms against viruses. The recently identified mammalian Type III IFN has biological effects similar to type I IFN. However, the biological effects of type III IFN have not yet been characterized in birds. We compared the effects of chicken type III IFN (IFN-λ) with type I (IFN-β) and type II (IFN-γ) IFNs on IFN-stimulated genes (ISGs) using recombinant proteins expressed in Escherichia coli. Recombinant chicken IFN-λ inhibited influenza virus replication and induced the mRNA expression of the ISGs, Mx and OAS, in chicken embryonic fibroblasts (CEFs) in a dose-dependent manner. However, the effective dose of IFN-λ was higher than that of IFN-β and IFN-γ. Furthermore, the effect of IFN-λ on induction of Mx and OAS was lesser than that of IFN-β, but comparable to that of IFN-γ. These results indicate that chicken IFN-λ has the potential to induce ISGs and inhibit viral replication in chicken cells.
TL;DR: Growth curves and reference values for hematological and serum biochemical parameters of Microminipigs showed no gender differences and the values were very similar to those in Göttingen and Yukatan minipigs.
Abstract: In this study, we demonstrated growth curves and reference values for hematological and serum biochemical parameters of Microminipigs, the world smallest experimental minipigs. In both male and female animals, the body weights (BWs) at 3 and 6 months of age were <5 kg and <10 kg, respectively, and growth curve revealed almost plateau (approximately 20 kg BW) after 18 months of age. Major hematological and serum biochemical parameters showed no gender differences and the values were very similar to those in Gottingen and Yukatan minipigs. The values obtained in this study can serve as fundamental reference, and thereby facilitate the use of Microminipig in life science research.
TL;DR: Factor affecting somatic cell counts in bovine bulk milk as a result of intramammary infections as well as non-infectious factors are presented and quality indicators in bulk tank milk are presented.
Abstract: The objective of this study was therefore to present factors affecting somatic cell counts in bovine bulk milk as a result of intramammary infections as well as non-infectious factors. The paper presents also the impact of on-farm management practices on the level of bulk milk somatic cell counts and presents quality indicators in bulk tank milk. At the farm level bulk milk bacterial infection takes place through three main sources: bacterial contamination from the external surface of the udder and teats, from the surface of the milking equipment, and from mastitis microorganisms within the udder. The threshold of 200,000 cells/ml identifies bacteriological negative quarters of the udder. The counts of mammary pathogens in bulk tank milk are relatively low, on average not exceeding 1,000 cfu/ml. Environmental pathogens predominate in bulk tank milk samples with somatic cells count <300 × 10(3) ml.
TL;DR: This study clarified the current status of the horses, obtained genetic information of individuals, and confirmed that the parentage test for identification using the 31 microsatellite DNAs was highly reliable, which increases the reliability of stud certification, and is also helpful for effective management.
Abstract: In order to contribute to conservation of the endangered Kiso horse, we clarified their genetic information using 31 microsatellite DNAs, and genotyped 125 horses, 83% of the existing breed. First, we clarified the current status of the horses. The horses were confirmed to have experienced rapid loss of population causing a bottleneck, and their effective population size was much smaller than their census size. Moreover, the number of alleles (6.3), observed heterozygosity (0.674), and expected heterozygosity (0.662) were in the same range as other endangered horses all over the world. Therefore, although their inbreeding level was not so severe (Fis: -0.017), the Kiso horse is surely one of the endangered. Second, we obtained genetic information of individuals. This information allowed us to understand the genetic distance of individuals, and might help in development of a reproductive strategy concerning the genetic distance between the mating pairs. Moreover, there appeared to be 4 subpopulations of Kiso horse, and this result was in good agreement with their historical background. Third, we confirmed that the parentage test for identification using the 31 microsatellite DNAs was highly reliable (probability of exclusion: 0.999999993). This identification increases the reliability of stud certification, and is also helpful for effective management. Understanding the genetic diversity within the population and the relationships among individuals is important to ensuring effective management for maintenance of genetic variation, and this study may help in conservation of the endangered Kiso horse.
TL;DR: In this article, the effect of propofol and thiopental on the plasma oxidant-antioxidant profile in dogs undergoing surgery at doses used to induce anesthesia was evaluated.
Abstract: The present study aimed to evaluate the effect of propofol and thiopental on the plasma oxidant-antioxidant profile in dogs undergoing surgery at doses used to induce anesthesia. The plasma total oxidant status (TOS) and oxidative stress index (OSI) levels increased significantly with time in both groups, whereas the plasma total antioxidant status (TAS) levels decreased with time in both groups. The OSI was significantly higher at the end of surgery than before induction of anesthesia in both groups. The TOS and OSI change ratio of propofol group were significantly lower than that of thiopental group. In conclusion, our findings show that propofol has antioxidant effects in dogs. Further studies need to be conducted to demonstrate the exact mechanism of oxidative stress due to anesthesia and surgery in dogs.
TL;DR: A significant negative correlation was observed between the lowest ruminal pH and its corresponding ruminal temperature in cows fed the C and RAI diets, suggesting active fermentation and volatile fatty acid production in the rumen.
Abstract: To assess the relationship between pH and temperature in the ruminal bottom fluid, circadian changes were monitored using cows fed a control diet (C diet) or a rumen acidosis-inducing diet (RAI diet) by using a wireless radio-transmission pH- measurement system. These two parameters were measured simultaneously at 10-min intervals on day 14 after commencement of feeding. Compared to the mean ruminal pH for 60 min immediately after the morning feeding (0 hr), significantly lower pH was noted 3-13 hr later (P<0.05) and 4-19 hr later (P<0.01) in cows fed the C and RAI diets, respectively, although the reduction in the latter was much higher than that in the former. In contrast, significantly higher ruminal temperature was found at 8 and 12-14 hr later (P<0.05) and 6, 8, and 10-19 hr later (P<0.01) in cows fed the C and RAI diets, respectively. A significant negative correlation was observed between the lowest ruminal pH and its corresponding ruminal temperature in cows fed the C and RAI diets (r=-0.722 and -0.650, P<0.01, respectively), suggesting active fermentation and volatile fatty acid production in the rumen. However, ruminal pH profiles may not be predictable by measuring only ruminal temperature because decreases in ruminal pH were preceded by increases in ruminal temperature, and circadian changes in pH and temperature were associated with ruminal fermentation.