Comparison of mark-resight methods to estimate abundance and rabies vaccination coverage of free-roaming dogs in two urban areas of south Bhutan.

In Bhutan, Capture-Neuter-Vaccinate-Release (CNVR) programs have been implemented to manage the dog population and control rabies, but no detailed evaluation has been done to assess their coverage and impact. We compared estimates of the dog population using three analytical methods: Lincoln-Petersen index, the Chapman estimate, and the logit-normal mixed effects model, and a varying number of count periods at different times of the day to recommend a protocol for applying the mark-resight framework to estimate free-roaming dog population abundance. We assessed the coverage of the CNVR program by estimating the proportion of dogs that were ear-notched and visually scored the health and skin condition of free-roaming dogs in Gelephu and Phuentsholing towns in south Bhutan, bordering India, in September-October 2012. The estimated free-roaming dog population in Gelephu using the Lincoln-Petersen index and Chapman estimates ranged from 612 to 672 and 614 to 671, respectively, while the logit-normal mixed effects model estimate based on the combined two count events was 641 (95% CI: 603-682). In Phuentsholing the Lincoln-Petersen index and Chapman estimates ranged from 525 to 583 and 524 to 582, respectively, while the logit-normal mixed effects model estimate based on the combined four count events was 555 (95% CI: 526-587). The total number of dogs counted was significantly associated with the time of day (AM versus PM; P=0.007), with a 17% improvement in dog sightings during the morning counting events. We recommend to conduct a morning marking followed by one count event the next morning and estimate population size by applying the Lincoln-Peterson corrected Chapman method or conduct two morning count events and apply the logit-normal mixed model to estimate population size. The estimated proportion of vaccinated free-roaming dogs was 56% (95% CI: 52-61%) and 58% (95% CI: 53-62%) in Gelephu and Phuentsholing, respectively. Given coverage in many neighbourhoods was below the recommended threshold of 70%, we recommend conducting an annual "mass dog vaccination only" campaign in southern Bhutan to create an immune buffer in this high rabies-risk area. The male-to-female dog ratio was 1.34:1 in Gelephu and 1.27:1 in Pheuntsholing. Population size estimates using mark-resight surveys has provided useful baseline data for understanding the population dynamics of dogs at the study sites. Mark-resight surveys provide useful information for designing and managing the logistics of dog vaccination or CNVR programs, assessing vaccination coverage, and for evaluating the impact of neutering programs on the size and structure of dog populations over time.

Japanese Black breeding herd exhibiting low blood urea nitrogen: a metabolic profile study examining the effect on reproductive performance.

Ten reared cows of a Japanese Black cattle herd in Kagoshima prefecture, Japan, exhibited extremely low blood urea nitrogen (BUN) concentration (2.6 ± 0.6 mg/dL). Examination of dietary feed nutrition and relevant pastureland soil content suggested a correlation with crude protein (CP) deficiency or unbalanced nutritional dietary feeds. Thirteen months after the introduction of a dietary remedial measure (bean cake supplementation), BUN, total cholesterol and albumin concentration from five of the original 10 cows increased significantly compared with their values of before the dietary remedy. The postpartum day open period was significantly lower after the dietary remedial measure than that before it. The abnormally low BUN levels of the cattle herd may be due to inadequate dietary nutritional content, primarily from the imbalance of total digestible nutrient and CP of the feed and far lower han average CP value. In conclusion, routine examination of serum biochemical parameters in Japanese Black breeding cattle may be a useful strategy for determining subclinical metabolic failure of cattle herds, and consequently, its effect on reproductive performance of the herd.

Effects of long-term in vitro exposure of ejaculated boar sperm to zearalenone and α-zearalenol in sperm liquid storage medium.

The effects of in vitro exposure of porcine spermatozoa to zearalenone (ZEN) and α-zearalenol (α-ZOL) were studied by evaluating several parameters of an in vitro fertilization (IVF) system. For this purpose, boar spermatozoa cultured with semen storage medium containing 0 (control), 10 and 1000 µg/L of ZEN and α-ZOL for 1 week at 5°C were used for IVF of in vitro matured oocytes. Overall, there were no significant differences in the rates of total penetration, monospermic fertilization, and polyspermic fertilization of oocytes inseminated with spermatozoa from the different groups. Similarly, ZEN and α-ZOL at 10 and 1000 µg/L did not have detrimental effects on the cleavage and development to blastocysts of oocytes after in vitro fertilization. Although the motility, viability, and plasma membrane integrity of spermatozoa significantly decreased after 3 weeks of storage compared to non-stored spermatozoa (P < 0.05), ZEN and α-ZOL at the evaluated concentrations did not exert detrimental effects on the above parameters, even after 3 weeks of storage. These results indicate that prolonged exposure of boar spermatozoa to ZEN and α-ZOL up to 1000 µg/L under reduced metabolic conditions does not affect their in vitro function.

Metabolic profile of Japanese Black breeding cattle herds: usefulness in selection for nutrient supplementation to enhance reproductive performance and regional differences.

The study aims were (1) to confirm the effects of nutritional improvement in prepartal and postpartal periods, monitored using the serum metabolic profile test (MPT) and reproductive performance, and (2) to clarify regional characteristics of the MPT results within our jurisdiction by using our MPT database. Experiment 1: Among 42 breeding cattle herds in our jurisdiction mainly fed home-pasture roughage, 3 experimental herds showing subnormal blood urea nitrogen (BUN) levels were selected and compared with 1 representative excellent herd. Dietary remedial measures were implemented from feed analysis in each herd. BUN concentration in all 3 herds increased significantly, and open days postpartum in 2 of the herds were significantly reduced, compared with values before dietary supplementation. Experiment 2: Thirty-seven herds within our jurisdiction were grouped into 3 categories (Area 1, 2 and 3) by location and soil condition of the herd pastureland. The MPT and reproductive performance in cows whose blood samples were collected at both prepartum (60-20 days before calving) and postpartum (30-90 days after calving) were compared among the 3 areas. Significant regional differences were found in prepartal albumin, total cholesterol, BUN, and glucose and postpartal BUN, glucose and open days (P<0.05). Overall, the MPT (especially BUN) might be useful for determining the metabolic nutritional status of breeding cattle herds, particularly those fed home-pasture roughage. Additionally, poor/unsatisfactory reproductive performance of beef breeding cattle herds probably reflects inadequate nutritional content of the diet, possibly arising from regional pastureland differences.

Detection of Torque teno sus virus 1 and 2 in tissues from stillborn piglets delivered by sows via natural farrowing.

We detected Torque teno sus virus 1 and 2 (TTSuV1 and TTSuV2) in tissue samples from 18 stillborn piglets using nested polymerase chain reaction. The detection rates of TTSuV1 and TTSuV2 were 78% and 50%, respectively, with 83% of the stillborn piglets positive for TTSuV1 or TTSuV2. TTSuV1 was detected highest in the liver (72%) followed by heart (56%), spleen (38%) and tonsils (38%) while TTSuV2 was detected highest in the tonsils (38%) followed by liver (33%), spleen (25%), and heart (17%). These results indicate that TTSuVs are commonly present but not equally distributed among the tissues of stillborn piglets.

Seroprevalence of Torque teno sus virus types 1 and 2 in postweaning multisystemic wasting syndrome-suspected pigs and porcine circovirus type 2-vaccinated normal pigs in southern Japan.

We investigated the seroprevalence rate of Torque teno sus virus types 1 (TTSuV1) and 2 (TTSuV2) in the sera of 38 post weaning multisystemic wasting syndrome (PMWS)-suspected pigs and 43 porcine circovirus type 2 (PCV2)-vaccinated normal pigs on 3 commercial pig farms in southern Japan by using nested polymerase chain reaction. High seroprevalence rate of TTSuVs was observed in both PMWS-suspected pigs (100%) and PCV2-vaccinated normal pigs (90.7%). The seroprevalence rate of TTSuV2 was significantly higher in the PMWS-suspected pigs than in PCV2-vaccinated normal pigs (97.4% versus 81.4%, P<0.05), whereas no such difference was observed for TTSuV1 between the 2 groups of pigs. In both pig groups, the seroprevalence rate of TTSuV2 was significantly higher than that of TTSuV1 (P<0.01-0.05). These results show that TTSuVs are highly seroprevalent in both PMWS-suspected pigs and PCV2-vaccinated normal pigs with TTSuV2 being more seroprevalent in the former than in the latter.

Detection of Torque teno sus virus types 1 and 2 by nested polymerase chain reaction in sera of sows at parturition and of their newborn piglets immediately after birth without suckling colostrum and at 24 hr after suckling colostrum.

This study was performed to clarify the sow-to-fetus transmission pathway of Torque teno sus virus (TTSuV) types 1 (TTSuV1) and 2 (TTSuV2). For this purpose, detection of TTSuV1 and TTSuV2 (TTSuVs) in sera of 6 sows (Sows 1-6) at parturition and in sera of their newborn piglets immediately after birth without suckling colostrum was performed by nested polymerase chain reaction (nPCR). These sows were bred using semen that had tested negative for TTSuVs. In a TTSuV1- and TTSuV2-positive sow (Sow 1), TTSuV1 and TTSuV2 were detected in 4 and 5 of 12 newborn littermates, respectively. In a TTSuV1-positive sow (Sow 2), TTSuV1 was detected in 1 of 8 newborn littermates. In 4 TTSuV1- and TTSuV2-negative sows (Sows 3-6), TTSuV1 was detected in 6 out of the 25 newborn piglets of 3 sows (Sows 3-5), while TTSuVs were not detected in all 13 piglets of 1 sow (Sow 6). In addition, to investigate the possibility of a sow-to-piglet transmission pathway of TTSuV via colostrum, TTSuV1 and TTSuV2 in sera of 12 newborn piglets from Sows 1-3 were examined by nPCR. Immediately after birth without suckling colostrum, TTSuV1 and TTSuV2 were not detected in 10 and 8 of 12 newborn piglets, respectively; however, at 24 hr after suckling colostrum, TTSuV1 was detected in 6 piglets, while TTSuV2 was not detected in any piglets. These results confirmed the existence of a sow-to-fetus transmission pathway of TTSuV during normal pregnancy and suggested a possibility of sow-to-piglet transmission of TTSuV via colostrum.

Infection dynamics of Torque teno sus virus types 1 and 2 in serum and peripheral blood mononuclear cells.

This study was carried out to investigate the presence of Torque teno sus virus types 1 (TTSuV1) and 2 (TTSuV2) in a longitudinally (14 to 150 days of age) collected paired pooled sera (pSE) and peripheral blood mononuclear cells (pPBMCs) using nested polymerase chain reaction. The detection rate of TTSuV1 in pSE increased from 14 to 90 days of age, but a progressive decline was observed from 120 to 150 days of age, while in pPBMC, a high value was maintained till the end of growing-finishing period. On the contrary, except in PBMCs at 30 days of age, high detection rates of TTSuV2 were found in both pSE and pPBMCs in all sampling ages. The detection rate of TTSuVs between pSE and pPBMCs was positively correlated at all sampling ages except for TTSuV1 at 150 days of age. This is the first study showing the presence of TTSuVs in PBMCs from pigs and describing the in vivo infection dynamics of TTSuV in paired sera and PBMCs during the entire growing and finishing periods of pigs reared in conventional farms.

Acute colic possibly caused by phytobezoar derived from napier grass in 5 Japanese black cows reared in the same farm.

For 6 years, 5 Japanese Black cows of the same herd showed anorexia, depression, and dehydration with no feces in the rectum. Biomedical examination of 3 animals showed severe hypokalemia and hypochloremia. Although the first 3 animals died or were slaughtered (causes unknown), necropsy results showed that the cow in case 4 had intestinal obstruction due to phytobezoar derived from napier grass, fed mainly to the cattle as roughage. Therefore, farmers were recommended to avoid the hard root-stem portion of napier grass as roughage. Consequently, less phytobezoar was recovered from the fifth cow, and no similar clinical case of intestinal obstruction was observed thereafter. This is the first report on intestinal obstruction caused by phytobezoars derived from napier grass.