Molecular characterization of tetracycline- and quinolone-resistant Aeromonas salmonicida isolated in Korea

The antibiotic resistance of 16 Aeromonas (A.) salmonicida strains isolated from diseased fish and environmental samples in Korea from 2006 to 2009 were investigated in this study. Tetracycline or quinolone resistance was observed in eight and 16 of the isolates, respectively, based on the measured minimal inhibitory concentrations. Among the tetracycline-resistant strains, seven of the isolates harbored tetA gene and one isolate harbored tetE gene. Additionally, quinolone-resistance determining regions (QRDRs) consisting of the gyrA and parC genes were amplified and sequenced. Among the quinolone-resistant A. salmonicida strains, 15 harbored point mutations in the gyrA codon 83 which were responsible for the corresponding amino acid substitutions of Ser⁸³→Arg⁸³ or Ser⁸³→Asn⁸³. We detected no point mutations in other QRDRs, such as gyrA codons 87 and 92, and parC codons 80 and 84. Genetic similarity was assessed via pulsed-field gel electrophoresis, and the results indicated high clonality among the Korean antibiotic-resistant strains of A. salmonicida.     

Improvement in the diagnosis of <Emphasis Type="Italic">Brucella abortus</Emphasis> infections in naturally infected water buffaloes (<Emphasis Type="Italic">Bubalus bubalis</Emphasis>) using an ELISA with a Protein-G-based indicator system

Brucellosis caused by Brucella abortus in domestic water buffaloes (Bubalus bubalis) raised under the traditional system of husbandry in northern India was diagnosed using an enzyme-linked immunosorbent assays (ELISA) with a Protein-G-based indicator system (Protein-G ELISA). A total of 1,551 animals that are positive (N = 61), negative (N = 243), and suspected (N = 1,247) for brucellosis were examined. Rose bengal test (RBT) was used to predict the disease, and accordingly, animals were dichotomized in positive and negative population for receiver operating characteristic (ROC) curve analysis to determine the sensitivity, the specificity, and the performance index of Protein-G ELISA. Taking all animals (N=1551) into account, the ROC curve analysis revealed cut off value of 29.6% positivity (%P) with 98.40% and 94.94%, sensitivity and specificity, respectively. The results were compared with ELISA in which anti-bovine conjugate was used. The cut off in ELISA was 37.9%P and sensitivity and specificity were 96.26% and 97.07%, respectively. The performance indexes of both the assays were almost equal and were 193.34 for Protein-G ELISA and 193.33 for ELISA. The cut off values of both the tests changed, if only known positive (N = 61) and known negative (N=243) animals were used for ROC curve analysis, and accordingly, changes in sensitivity and specificity were observed with significant decrease of performance indexes of both the tests. The high optical density (P<0.0001) background signal with negative serum control and high %P (P<0.0001) in sera from negative population were noticed in ELISA in comparison to Protein-G ELISA.     

In vitro inhibitory effect of trichostatin A on canine grade 3 mast cell tumor

Mast cell tumor (MCT) is one of the most prevalent neoplasms that affect skin and soft tissue in dogs. Because mast cell tumors present a great variety of clinical appearance and behavior, their treatment becomes a challenge. Trichostatin A (TSA), an antifungal antibiotic, has shown inhibitory effects on the proliferation and induction of apoptosis in various types of cancer cells. In order to evaluate the potential of trichostatin A as a therapeutic drug, cells of grade 3 MCT were cultured and treated with concentrations of 1 nM to 400 nM of TSA. MTT assay and trypan blue exclusion assays were performed to estimate cell growth and cell viability, and cell cycle analysis was evaluated. TSA treatment showed a reduction in numbers of viable cells and an increase of cell death by apoptosis. The cell cycle analysis showed an increase of hypodiploid cells and a reduction of G0/G1 and G2/M –phases. According to these results, trichostatin A may be an interesting potential chemotherapeutic agent for the treatment of canine MCT.     

Characterization of Doayo and Kapsiki taurine cattle breeds of Cameroon in their natural environment

Data were collected on live weight (LW), heart girth (HG), height-at-withers (HW), trunk length (TL), age, sex, and coat color of 207 taurines cattle—122 of the Doayo (Namchi) breed and 85 of the Kapsiki (Kirdi) breed. The animals, aged 1 to 20 years, were selected from 60 herds randomly selected from villages of Poli of Faro and Mokolo of Tsanga, divisions of the North and Far North Regions of Cameroon. The data were analyzed using the SAS program with a linear model, applying standard tests. Results indicated no breed effect (P > 0.05) in the growth trends of LW, HG, HW, and TL. HG and TL were highly significantly (P < 0.0001) related to LW. The growth pattern for the two breeds was the same since the linear contrast of least square means for the traits at various age groups did not differ (P > 0.05) significantly. The breeds attained maturity as from 4 years. In the absence of breed effect (P > 0.05), a single regression equation was established for the estimation of live weight as thus LW = - 244.42 ( ±22.57 ) kg + 2.49 ( ±0.23 ) HG + 1.04 ( ±0.25 ) TL {\hbox{LW}} = - {244}.{42 }\left( {\pm {22}.{57}} \right){\hbox{ kg}} + {2}.{49 }\left( {\pm 0.{23}} \right){\hbox{ HG}} + {1}.0{4 }\left( {\pm 0.{25}} \right){\hbox{ TL}} , with HG contributing up to 70% of total variation and TL, 2%. This equation could be used to develop a measuring band useful in the rural environment for commercial and clinical veterinary purposes.     

Effects of surgical implantation of temperature dataloggers on reproduction of captive striped skunks (Mephitis mephitis).

A total of 20 (14 females, six males) captive striped skunks (Mephitis mephitis) with miniature temperature dataloggers implanted free in the abdominal cavity were examined for reproductive performance and pathology because of implants. Eleven of 12 female skunks reproduced successfully 45.9+/-3.7 days after surgery to remove implanted dataloggers. The pregnancy rate of 91.7% (11/12) was much higher than other captive studies and was comparable with that of wild skunks. Eight striped skunks (six males, two females) that were euthanatized and necropsied after having implants in the abdominal cavity for 5 mo showed no apparent pathology associated with the implant. Implantation of the intraperitoneal devices did not lead to complications in abdominal tissues. Neither implant nor surgery affected reproduction. We conclude that implanted dataloggers can safely be used in physiologic studies of striped skunks or possibly other small carnivores in captive or field studies.     

Effect of muscle type and vacuum chiller ageing period on the chemical compositions,meat quality,sensory attributes and volatile compounds of Korean native cattle beef

The present study demonstrates the effects of different muscle types and chiller ageing periods on the chemical composition, meat quality parameters, sensory characteristics and volatile compounds of Karean native cattle beed. Longissimus dorsi (LD) and Semitendinosus (ST) muscles aged for 7 days and 28 days were used. Moisture, cooking loss, total collagen and Warner‐Bratzler shear force (WBSF) values for the ST were higher than the LD muscle regardless of ageing period (P < 0.05). The LD muscle had higher intramuscular fat (IMF) (P < 0.05). Ageing for 28 days decreased WBSF values whereas it increased thiobarbituric acid of both muscles. Moreover, tenderness, juiciness and flavor scores were significantly higher for the LD muscle at both ageing periods. Increased ageing time improved tenderness of both muscles, and increased juiciness of the LD muscle, whereas there was decreased flavor score of ST muscle (P < 0.05). The majority of the volatile compounds formed from the oxidation of lipids showed differences between the two muscles. Ageing for 28 days increased in the amounts of many volatile compounds; however, the amounts of some important volatile compounds were decreased. These results clearly demonstrate that muscle type and ageing have a potential effect on meat quality, sensory characteristics and volatile profile.     

Growth,feed efficiency and blood profile of buffalo calves consuming high levels of fluoride

Twelve male buffalo calves of 10 to 12 months of age were divided into 3 groups of four each. They were fed wheat straw+concentrate mixture +3 Kg greens. The chemical composition of the diet was same in all the three groups except fluoride which was added (as NaF) in concentrate mixture of group B and C to make the final fluoride concentration 30 ppm and 60 ppm respectively. The animals were kept on scheduled diet for a period of 90 days. Body weights were recorded at the start of the experiment and at fortnightly interval thereafter. Analysis of data revealed that the dry matter intake decreased non significantly in group B and C as compared to control group. A significant decrease in serum calcium and a significant increase in phosphorus concentration were observed in group C animals. A significant increase was observed in alkaline phosphatase activity in group C animals. A non significant decrease was observed in T₄ values in group C animals. On the basis of these results it could be concluded that fluoride in the diet of buffalo calves @ 30 ppm is a safe level whereas 60 ppm has affected the blood metabolites.     

Production of blastocysts after intergeneric nuclear transfer of goral (Naemorhedus goral) somatic cells into bovine oocytes

Interspecies cloning may be a useful method to help conserve endangered species and to study nuclear-cytoplasm interaction. The present study investigated in vitro development of goral (Naemorhedus goral) intergeneric nuclear transfer embryos produced by fusing goral fibroblasts with enucleated metaphase II (MII) bovine oocytes. After two to five passages, serum-starved or non-starved goral skin fibroblast cells were transferred into enucleated MII bovine oocytes. Couplets were electrically fused and chemically activated, and then cultured in either modified synthetic oviduct fluid (mSOF) or tissue culture medium-199 (TCM-199) supplemented with 10% FBS. Serum starvation of donor cells did not affect the fusion rate and or development to of cells to the two-cell stage, to more than 9-cells, or to morulae, regardless of culture medium. Three blastocysts from 202 fused embryos were obtained when embryos reconstructed with non- serum- starved donor cells were cultured in mSOF. However, no blastocysts were obtained when the embryos reconstructed with serum-starved donor cells were cultured in mSOF. The total cell number of goral intergeneric embryos averaged 130.3 (range 105-180). In conclusion, this study demonstrated that bovine oocytes can support blastocyst development after intergeneric SCNT with goral fibroblasts.     

Dynamics of natural bovine herpesvirus-1 (BoHV-1) infection in a dairy herd

In this study, natural cycling of BoHV-1 infection was investigated in two groups of dairy cattle containing 2120 head. Group 1 comprised 127 animals and they were monitored for BoHV-1 infection virologically and serologically in six consecutive sampling periods. It consisted of naive heifers between 6 and 8 months of age, while in group 2, age, sex and the BoHV-1 serostatus of the animals were disregarded. The animals in group 1 were found to have seroconverted at the second sampling. Results of the serological study showed slight antibody response after natural BoHV-1 infection in the herd and neutralizing titres fell below protective levels in the 6–8 months after the peak. During the 2-year study period, one recurrence was detected after primary infection. Virus isolation studies revealed a cytopathic effect indicative of BoHV-1 in two nasal swabs taken during the fifth sampling period from animals with mild upper respiratory tract symptoms. As the study was carried out under natural conditions, it is not known whether the viruses isolated were from recurrences or re-infections. Data from cross-neutralization tests with herd isolates showed higher antibody response than those with the reference virus. The dynamics of BoHV-1 in both groups were found to be statistically similar.     

Frequent use of colistin-based drug treatment to eliminate extended-spectrum beta-lactamase-producing <Emphasis Type="Italic">Escherichia coli</Emphasis> in backyard chicken farms in Thai Binh Province,Vietnam

Reports of livestock infections with extended-spectrum beta-lactamase-producing Escherichia coli (ESBL-E) are increasing. Based on interviews conducted over a 6-month period, we found that veterinarians in the Vietnamese province of Thai Binh prefer to prescribe colistin-based drugs (CBD) in chicken farms. We aimed to clarify whether CBD use selects for strains of colistin-resistant ESBL-E. With the cooperation of seven local households, we detected ESBL-E in chickens’ feces after treating chickens with CBD. Phylogenetic groupings and the presence of CTX-M/AmpC genes were determined, and the multi-antibiotic susceptibility of isolates was analyzed. Our results showed that ESBL-E presented in seven chickens’ feces from two households. Seventy-two percent of ESBL-E isolates harbored CTX-M9 and the phylogenetic group A; the colistin minimum inhibitory concentration (MIC) of all isolated ESBL-E ranged from 0.064 to 1 μg mL^?1. Moreover, ESBL-E isolates were used to experimentally select for colistin resistance, and the effect of commercial CBD on ESBL-E was investigated. The results showed that an ESBL-E strain with a colistin MIC of 4 μg mL^?1 was able to grow in media with CBD. Although CBD treatment was effective, in vitro experiments demonstrated that ESBL-E can easily acquire colistin resistance. Therefore, restrictions on colistin use are necessary to prevent the emergence of colistin-resistant bacteria.