Livestock movement patterns in the main livestock production provinces of Lao PDR.

Foot-and-mouth disease (FMD) is a highly infectious transboundary disease that is endemic and affects the livelihood of smallholder farmers in Lao People's Democratic Republic (PDR). Knowledge about livestock movement patterns is important for preventing the spread of FMD between villages. This study describes the livestock movement patterns in Champasak, Savannakhet and Xiangkhouang provinces of Lao PDR. Face-to-face interviews were conducted with randomly selected villagers (n = 195) and traders (n = 169) in 115 villages between February and March 2019. Livestock owners commonly purchased (mainly breeding) animals from other smallholders (81%) and sold (mainly slaughter) animals to traders (76%) or other smallholders (16%), typically within the same district and province. The median inter-village trade distance was 20-30 km, with an average frequency of 4 trades per village per month. Traders purchased animals from smallholders (71%) and middlemen (25%) located within their district. It was common for many traders (74%) to retain animals at their property before selling, typically a median of 4 beef cattle per trader. Local trades within the district were far more common (72%) than distant trades. The movements of grazing/fattening large ruminants between villages were reported in 30% of the villages in all three provinces and occurred mostly within the same district or province in short distance (6 km). Social Network Analysis has identified animal movement hubs in the three provinces which could be targeted for FMD control and surveillance. Movements of animals for further use (fattening/ reproduction), long-distance movements and frequent local movements described in this area have important implications for FMD circulation. The findings from the study will inform FMD spread simulation models for Lao PDR. The knowledge gained from these data will also help the Lao PDR authorities understand the patterns of animal movements associated with disease spread.

Longitudinal serological survey and herd-level risk factors for Leptospira spp. serovars Hardjo-bovis and Pomona on deer farms with sheep and/or beef cattle.

AIM: To investigate the seroprevalence of Leptospira spp. serovars Hardjo-bovis and Pomona on deer and mixed deer, sheep and/or beef cattle farms in the lower North Island of New Zealand and to examine associations between putative risk factors for seropositive deer herds. METHODS: Serological screening was conducted on 19 commercial deer farms, 16 with sheep and/or beef cattle, between August and October each year between 2006 and 2008. No leptospiral vaccination had been conducted on the farms. On each farm every year, serum samples were collected from a random sample of 20 or more rising 2-year-old replacement animals from each species. The microscopic agglutination test (MAT) was used to detect leptospiral antibodies against Leptospira borgpetersenii serovar Hardjo-bovis and Leptospira interrogans serovar Pomona. For both serovars, a titre of ≥1:48 was considered positive and a herd was considered seropositive if >3 of 20 serum samples were positive. Information on potential herd-level risk factors for deer herds being seropositive was obtained from a questionnaire completed by the farm owner or manager. RESULTS: The mean percentage of deer, cattle and sheep herds seropositive for Hardjo-bovis alone over 3 years was 42%, 53% and 54%, respectively, and for serovar Pomona was 7%, 5% and 0%, respectively. Antibodies to both serovars were found in 23%, 16% and 10% of deer, cattle and sheep herds, respectively. At the individual animal level, 228/1,107 (21%) deer, 308/767 (40%) cattle and 369/1,244 (30%) sheep were seropositive for Hardjo-bovis, 102 (9%) deer, 51 (7%) cattle and 23 (2%) sheep were seropositive for Pomona, and 49 (4%) deer, 28 (4%) cattle and 18 (1%) sheep were seropositive for both serovars. Deer herds were more likely to be seropositive for Hardjo-bovis in 2006 than 2008 (p=0.008), when seropositive in the preceding year (p=0.016) and on hilly compared with flat topography (p<0.001). Deer herds were more likely to be seropositive for Pomona when seropositive in the preceding year (p=0.016), when co-grazing with sheep flocks that were seropositive for Pomona (p<0.001), and when herds had a closed- compared with open-herd replacement policy (p<0.001). CONCLUSIONS: Exposure to Leptospira spp. was widely distributed in deer, sheep and beef cattle in the lower North Island of New Zealand. Co-grazing of deer with sheep that were seropositive was a potential risk factor for deer herds to be seropositive.

Growth response and shedding of Leptospira spp. in urine following vaccination for leptospirosis in young farmed deer.

AIM: To investigate the effect of vaccination against Leptospira serovars Hardjo-bovis and Pomona on growth rate and shedding of leptospires in urine in rising 1-year-old farmed red deer. METHODS: In early March 2007, 230 female and 205 male, 3-month-old deer on five farms were treated with streptomycin then were randomly allocated to a control group (n = 218), or were vaccinated (n = 217) with a bivalent whole-cell killed leptospiral vaccine (Leptavoid-2) followed by a booster 4 weeks later. These animals were isolated from other 3-month-old deer on each property until May, when all vaccinated and control deer were combined with deer not treated with streptomycin, for maximum exposure to natural leptospiral challenge. Seroconversion was monitored in control deer against Leptospira borgpetersenii serovar Hardjo-bovis and Leptospira interrogans serovar Pomona using the microscopic agglutination test (MAT). A cut-off of ≥ 1:48 was used to identify a seropositive response to infection, and ≥1:24 to indicate a response to vaccination. Urine was collected from females in all groups and tested for shedding of leptospires using bacterial culture and real-time PCR. Male deer from vaccinated and control groups were weighed every 2-3 months to measure growth rate. RESULTS: In vaccinated female deer, antibody was present to Hardjo-bovis in 39-73% and to Pomona in 78-100% of animals in May, with titres ranging between 1:24-1:96 and 1:24-1:1536, respectively; control female deer were all serologically negative. In November, female control deer were seropositive to Hardjo-bovis on 4/5 farms (seroprevalence range 4-78%) but seronegative to Pomona on all farms. Between March and November mean growth rate was higher in vaccinated than control deer (26 g/day) on one farm (p = 0.049) that had the highest prevalence of seroconversion (78-88%) and urinary shedding (56%). Between August and November on 4/5 farms, when almost all seroconversions occurred, growth rates were higher in vaccinated than control deer (31 g/day; p = 0.011). Urinary shedding of leptospires was detected by culture and/or PCR in 8/34 (24%) unvaccinated control and 20/38 (53%) animals not treated with streptomycin on 2/5 farms; no shedding was detected in vaccinated deer. The average risk difference for shedding between vaccinated and control deer was -11% (95%CI = -18 to -4) (p = 0.001). CONCLUSION: Vaccination against Leptospira serovars Hardjo-bovis and Pomona was associated with higher liveweight gain and reduced urinary shedding of leptospires in young deer naturally challenged by Hardjo-bovis. The association with liveweight gain was evident on some but not all farms.

Vaccination for leptospirosis improved the weaning percentage of 2-year-old farmed red deer hinds in New Zealand.

AIM: To investigate the effect of leptospiral vaccination against serovars Hardjo-bovis and Pomona on fetal loss and weaning percentage in rising 2-year-old farmed red deer hinds. METHODS: In mid-February 2007, 252 rising 2-year-old hinds on four farms received a single dose of streptomycin (25 mg/kg), to minimise leptospiral infection. They were randomly allocated to vaccinated and control groups. Vaccinated hinds (n=125) received a 2-ml S/C injection of a bivalent whole-cell killed leptospiral vaccine (Leptavoid-2) followed by a booster 4-6 weeks later, and were grazed with control hinds (n=127). These animals were isolated from other hinds on each property, until after mating (June 2007), when all vaccinated and control hinds were combined with hinds not treated with streptomycin, for maximum exposure to natural leptospiral challenge. Evidence of natural challenge by Leptospira spp. was assessed in blood samples from control hinds by serology against L. borgpetersenii serovar Hardjo-bovis and L. interrogans serovar Pomona, using the microscopic agglutination test (MAT), and in hinds not treated with streptomycin by detection of shedding of organisms in urine, using bacterial culture and real-time PCR. Pregnancy diagnosis was carried out in May/June 2007, using transrectal ultrasonography, to determine conception. In late October, prior to calving, the pregnant vaccinated and control hinds were examined by palpation of the abdomen and udder, to determine the percentage of hinds pregnant at term and assess fetal loss. In March 2008, at weaning, vaccinated and control hinds were examined for lactation status, using observation and palpation of the udder. The differences between the groups were evaluated using matched logistic regression analysis. RESULTS: After mating, pregnancy was diagnosed in 97/125 (77.6%) vaccinated and 106/127 (83.5%) control hinds. All four farms had serological evidence of Hardjo-bovis infection, and a single hind was serologically positive for Pomona between October and March. Real-time PCR confirmed urinary shedding on two farms. The mean percentage of hinds pregnant at term, for those animals confirmed pregnant after mating, in the vaccinated and control groups was 95/97 (98%) (range 95-100%) and 103/106 (97%) (range 94-100%), respectively (p>0.05). The mean weaning percentage for vaccinated and control groups was 86/97 (89%) (range 78-95%) and 88/106 (83%) (range 76-88%), respectively (p=0.015). CONCLUSION: Vaccination for leptospirosis resulted in no difference in the percentage of hinds pregnant at term, but a higher weaning percentage compared with unvaccinated controls suggesting that vaccination reduced perinatal and/or pre-weaning mortality.

Serosurvey of leptospirosis and investigation of a possible novel serovar Arborea in farmed deer in New Zealand.

AIM: To investigate the prevalence of Leptospira spp. and possible novel serovar Arborea infection in farmed deer in New Zealand. METHODS: In September 2006, five serum samples from a serum bank from each of 70 farms sampled for a previous national prevalence survey were forwarded to the World Health Organisation/Food and Agriculture Organisation/World Organisation for Animal Health (WHO/FAO/OIE) reference laboratory for leptospirosis in Brisbane, Australia, to test for reactivity to a reference panel of 23 serovars, most believed to be exotic to New Zealand, using the microscopic agglutination test (MAT). Eleven farms were seropositive for Arborea, a serovar novel to New Zealand. In July 2007, 126 additional banked serum samples from nine of those 11 farms (n=8-20/farm) were sent to the reference laboratory for similar serology. Two farms in the Southland region were considered positive for serovar Arborea. Tissue from deer kidneys (n=43) from these two farms collected at a deer slaughter premises (DSP) was cultured in November 2007 and November 2008. Sera from those deer were also sent to the laboratory in Brisbane. RESULTS: From the initial 350 sera, 96 (27.4%) and 19 (5.4%) samples were positive for Leptospira borgpetersenii serovar Hardjo-bovis and Leptospira interrogans serovar Pomona respectively. There were cross-reactions between serovar Hardjo-bovis with serovars Medanensis and Szwajizak. Serological evidence of serovars Tarassovi, Grippotyphosa, Celledoni, Australis, Zanoni, Robinsoni, Canicola, Kremastos, Bulgarica, Cynopteri, Ballum, Bataviae, Djasiman, Javanica, Panama, Shermani and Topaz was negative or sporadic, generally with titres of 1:50 and therefore likely non-specific. Fourteen (4.0%) samples from 11 farms were positive for serovar Arborea, justifying further investigation. The prevalence of serovar Arborea was 15% and 30% on two farms, from the 126 samples. None of 43 kidney and serum samples collected subsequently from those two farms were positive by culture or serology for serovar Arborea. CONCLUSIONS: While there were samples serologically positive for serovar Arborea in deer, attempts to isolate the organism were unsuccessful. The sample size for the follow-up investigation was insufficient to validate the presence or absence of infection, so further study should be undertaken to verify the status of this serovar of Leptospira spp. in New Zealand, in both deer and other livestock species.

Investigation of localisation of Leptospira spp. in uterine and fetal tissues of non-pregnant and pregnant farmed deer.

AIM: To find evidence for localisation in the uterus, and fetal infection, of Leptospira spp. in farmed deer in the lower North Island of New Zealand during and shortly after the breeding season. METHODS: Between February and July 2008, 116 blood samples, 120 kidneys, 120 uteri and 27 fetuses were collected from 120 mixed-age hinds from lines from nine farms, at a deer slaughter premises. Serum samples were tested for antibodies against Leptospira borgpetersenii serovar Hardjo-bovis and Leptospira interrogans serovar Pomona, using the microscopic agglutination test (MAT). For both serovars, a titre of ≥1:48 was considered positive. Samples from kidneys, uteri and fetal tissue were subjected to bacterial culture, using Ellinghausen-McCullough-Johnson-Harris (EMJH) medium, and real-time PCR, using DNA gyrase subunit B gene primers. RESULTS: Thirty-four of 116 (29.3%) serum samples were positive for serovar Hardjo-bovis, and 13 (11.2%) for serovar Pomona. Seven of 120 kidneys were positive for serovar Hardjo-bovis by culture, and five of these, but no others, were positive by real-time PCR. Of 120 uteri, none was culture- or PCR-positive. None of 27 fetal samples was culture-positive but one was positive by real-time PCR. The dam of the PCR-positive fetus was culture-negative from the kidney, but had an MAT titre of 1:192 for Hardjo-bovis. CONCLUSIONS: Attempts to isolate Leptospira spp. from the genital tracts and early fetuses of farmed deer were unsuccessful. However, molecular evidence suggested fetal infection in one case. This finding justifies further study of the role of leptospires in the genital tract and fetus and its association with reproductive loss in farmed deer.