Propagation of Asian isolates of canine distemper virus (CDV) in hamster cell lines

Backgrounds

The aim of this study was to confirm the propagation of various canine distemper viruses (CDV) in hamster cell lines of HmLu and BHK, since only a little is known about the possibility of propagation of CDV in rodent cells irrespective of their epidemiological importance.

Methods

The growth of CDV in hamster cell lines was monitored by titration using Vero.dogSLAMtag (Vero-DST) cells that had been proven to be susceptible to almost all field isolates of CDV, with the preparations of cell-free and cell-associated virus from the cultures infected with recent Asian isolates of CDV (13 strains) and by observing the development of cytopathic effect (CPE) in infected cultures of hamster cell lines.

Results

Eleven of 13 strains grew in HmLu cells, and 12 of 13 strains grew in BHK cells with apparent CPE of cell fusion in the late stage of infection. Two strains and a strain of Asia 1 group could not grow in HmLu cells and BHK cells, respectively.

Conclusion

The present study demonstrates at the first time that hamster cell lines can propagate the majority of Asian field isolates of CDV. The usage of two hamster cell lines suggested to be useful to characterize the field isolates biologically.     

Molecular epidemiology of Newcastle disease viruses in Vietnam

Newcastle disease virus (NDV) causes significant economic losses to the poultry industry in Southeast Asia. In the present study, 12 field isolates of NDV were recovered from dead village chickens in Vietnam between 2007 and 2012, and were characterized. All the field isolates were classified as velogenic. Based on the sequence analysis of the F variable region, two distinct genetic groups (Vietnam genetic groups G1 and G2) were recognized. Phylogenetic analysis revealed that all the 12 field isolates fell into the class II genotype VII cluster. Ten of the field isolates, classified as Vietnam genetic group G1, were closely related to VIIh viruses that had been isolated from Indonesia, Malaysia, and Cambodia since the mid-2000s, while the other two field isolates, of Vietnam genetic group G2, clustered with VIId viruses, which were predominantly circulating in China and Far East Asia. Our results indicate that genotype VII viruses, especially VIIh viruses, are predominantly responsible for the recent epizootic of the disease in Vietnam.     

Mapping the pork value chain in Vietnam: a systematic review

Tropical Animal Health and Production - In Vietnam, pork is the most commonly consumed type of meat, and the demand is expected to rise even further. Nevertheless, food safety is a major concern,...     

Aflatoxin B<Subscript>1</Subscript> (AFB<Subscript>1</Subscript>) reduces growth performance,physiological response,and disease resistance in Tra catfish (<Emphasis Type="Italic">Pangasius hypophthalmus</Emphasis>)

Triplicate groups of one hundred Tra catfish (8 g?±?0.2) were fed seven test diets containing increasing levels of AFB₁ (0, 50, 100, 250, 500, and 1000 μg AFB₁ kg^?1). Additionally Mycofix® Secure was added at 1.5% to one diet containing 500 μg AFB₁ kg^?1. Results showed that Tra catfish are sensitive to AFB₁. Reduction in weight gain (P?<?0.05) was observed for fish fed 50 μg AFB₁ kg^?1 and declined further with increasing levels of AFB₁ in the diets. Fish fed diets contaminated with 500 and 1000 μg AFB₁ kg^?1 showed increased (P?>?0.05) hepatosomatic index (HIS), while an increase in adipose somatic index (ASI) was observed in fish fed 50 μg AFB₁ kg^?1 and above when compared to the control and Mycofix® diets. After 12 weeks, blood serum analysis revealed higher alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in fish fed the 50, 100, and 250 μg AFB₁ kg^?1 suggesting occurrence of liver damage. Disease resistance of fish exposed to Edwardsiella ictaluri was also compromised by the presence of AFB₁ in the feed and was directly related to the contamination level. Seven days after Edwardsiella ictaluri exposure, survival rates were 50, 41.7, 31.7, and 8.3% for fish fed control, 50, 100, and 250 μg AFB₁ kg^?1, respectively. This trial shows that AFB₁ at a level of 50 μg AFB₁ kg^?1 and above can affect fish performance and disease resistance. Application of an effective mycotoxin management in the feed seems to be useful to prevent the negative effects of AFB₁.     

Relationship between total transit time and faecal quality in adult dogs differing in body size

Fed the same diet, large and giant-breed dogs have higher faecal moisture and increased frequency of soft stools than small ones. This could be the result of physiological differences, such as a different gastrointestinal transit time. In this study, we have correlated mean total transit time (MTT) with body size and faecal consistency in dogs varying in body size. Fifty dogs from 13 different breeds were used, from a Dachshund to a Great Dane. The MTT was determined using coloured plastic beads [Cummings and Wiggins, Gut, Vol. 17 (1976), p. 219], and faecal consistency was scored daily during the study. We confirmed the strong correlation between height at the shoulder (body size) and faecal score (r = 0.76; p < 0.0001). The MTT increased with body size, from 22 h for a Miniature Poodle to 59 h for a Giant Schnauzer. We found significant positive correlations (p < 0.0001) between MTT and body size as well as faecal scores (r = 0.71 and 0.70 respectively). In the present study, we observed an effect of body size on MTT. In our 50 healthy dogs a longer MTT was related to a poorer faecal quality. Previous studies reported no relationship between body size and the upper gastrointestinal transit time in healthy dogs. So, we hypothesized that body size would mainly affect colonic transit time and that a longer colonic residence time would be related to a poorer faecal quality by promoting fermentation activity.     

One year's study of dynamic and evolution of types I and II PRRSV in a swine farm

This study was to investigate dynamic and evolution of PRRSV in a seed-stock farm by monitoring PRRSV status from 11 June 2009 to 4 August 2010. For laboratory test, around 18-24 umbilical cords from farrowed sows and 5-95 sera from nursery and grow/finish pigs were submitted around every 2 weeks interval during the study. The submitted samples were tested for PRRSV using IDEXX PRRS 2XR ELISA kit, RT-nested PCR. The PRRSV-positive samples were further sequences based on ORF5 and analyzed using MEGA 3.1 program and Beast 1.5.4 package. The surveyed farm was first infected with type II PRRSV but it was infected newly with type I PRRSV of unknown origin, showing rapid substitution to type I PRRSV as a dominant strain in 2 weeks. The type I PRRSV was first detected from umbilical cord of a farrowed sow in 12 January 2010, and secondly from nursery pigs in 26 January 2010. Although sudden increase of mean S/P ratio was found in grow/finish pigs around 2 months earlier than first type I PRRSV detection, no type I PRRSV viremia was found. Thirty three ORF5 full sequences from 14 type II to 19 type I PRRSVs were obtained chronologically in this farm and the genetic characteristics and evolution rates of those sequences were analyzed. The substitution rates (/site/day) of two types were 4.03×10(-5) (type I), 3.09×10(-5) (type II), respectively, which was more frequent than previous reports. The calculated divergence time of type I PRRSV was consistent with the time when the sudden elevation of serum IgG in grow/finish barn was first observed. This study provided fundamental data for type I PRRSV dynamic in a previously type II PRRSV-infected farm and suggested grow/finisher barn could be a primary site for PRRSV introduction.     

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.     

Nuclear Replacement of In Vitro‐Matured Porcine Oocytes by a Serial Centrifugation and Fusion Method

The objective of the present study was to establish a method for nuclear replacement in metaphase‐II (M‐II) stage porcine oocytes. Karyoplasts containing M‐II chromosomes (K) and cytoplasts without chromosomes (C) were produced from in vitro‐matured oocytes by a serial centrifugation method. The oocytes were then reconstructed by fusion of one karyoplast with 1, 2, 3 or 4 cytoplasts (K + 1C, K + 2C, K + 3C and K + 4C, respectively). Reconstructed oocytes, karyoplasts without fusion of any cytoplast (K) and zona‐free M‐II oocytes (control) were used for experiments. The rates of female pronucleus formation after parthenogenetic activation in all groups of reconstructed oocytes (58.2–77.4%) were not different from those of the K and control groups (58.2% and 66.0%, respectively). In vitro fertilization was carried out to assay the fertilization ability and subsequent embryonic development of the reconstructed oocytes. The cytoplast : karyoplast ratio did not affect the fertilization status (penetration and male pronuclear formation rates) of the oocytes. A significantly high monospermy rate was found in K oocytes (p < 0.05, 61.6%) compared with the other groups (18.2–32.8%). Blastocyst formation rates increased significantly as the number of the cytoplasts fused with karyoplasts increased (p < 0.05, 0.0–15.3%). The blastocyst rate in the K + 4C group (15.3%) was comparable with that of the control (17.8%). Total cell numbers in both the K + 3C and K + 4C groups (16.0 and 15.3 cells, respectively) were comparable with that of the control (26.2 cells). Our results demonstrate that a serial centrifugation and fusion (Centri‐Fusion) is an effective method for producing M‐II chromosome transferred oocytes with normal fertilization ability and in vitro development. It is suggested that the number of cytoplasts fused with a karyoplast plays a critical role in embryonic development.