Epidemiological Survey and Risk Factor Analysis of 14 Potential Pathogens in Golden Snub-Nosed Monkeys at Shennongjia National Nature Reserve, China.

Golden snub-nosed monkeys (Rhinopithecus roxellanae) belong to Class A, the highest level of endangered primate species. Exploring the infection status of potential pathogens in golden snub-nosed monkeys is important for controlling associated diseases and protecting this species. The objective of this study was to investigate the seroprevalence for a number of potential pathogens and the prevalence of fecal adenovirus and rotavirus. A total of 283 fecal samples were collected from 100 golden snub-nosed monkeys in December 2014, June 2015, and January 2016; 26 blood samples were collected from 26 monkeys in June 2014, June 2015, January 2016 and November 2016 at Shennongjia National Reserve in Hubei, China. The infection of 11 potential viral diseases was examined serologically using an Indirect Enzyme-linked Immunosorbent Assay (iELISA) and Dot Immunobinding Assays (DIA), while the whole blood IFN-γ in vitro release assay was used to test tuberculosis (TB). In addition, fecal Adenovirus and Rotavirus were detected using Polymerase Chain Reaction (PCR). As a result, the Macacine herpesvirus-1 (MaHV-1), Golden snub-nosed monkey cytomegalovirus (GsmCMV), Simian foamy virus (SFV) and Hepatitis A virus (HAV) were detected with the seroprevalence of 57.7% (95% CI: 36.9, 76.6), 38.5% (95% CI: 20.2, 59.4), 26.9% (95% CI: 11.6, 47.8), and 7.7% (95% CI: 0.0, 84.2), respectively. Two fecal samples tested positive for Adenovirus (ADV) by PCR, with a prevalence of 0.7% (95% CI: 0.2, 2.5), and further, the amplification products were sequenced. Phylogenetic analysis revealed that they belonged to the HADV-G group. However, other pathogens, such as Coxsackievirus (CV), Measles virus (MeV), Rotavirus (RV), Simian immunodeficiency virus (SIV), Simian type D retroviruses (SRV), Simian-T-cell lymphotropic virus type 1 (STLV-1), Simian varicella virus (SVV), Simian virus 40 (SV40) and Mycobacterium tuberculosis complex (TB) were negative in all samples. In addition, a risk factor analysis indicated that the seroprevalence of MaHV-1 infection was significantly associated with old age (≥4 years). These results have important implications for understanding the health status and conservation of the endangered golden snub-nosed monkey population at Shennongjia Nature Reserve.

An outbreak of tuberculosis in endangered northern pig-tailed macaques (Macaca leonina) and milu deer (Elaphurus davidianus) from a zoo in China.

BACKGROUND: Tuberculosis (TB) is a zoonotic disease that affects humans and domesticated and wild animals. Animals in zoos are potentially an important source of TB for humans; however they are often neglected in routine disease surveillance programs. This investigation reports an outbreak of TB in milu deer and northern pig-tailed macaques in a zoo in Wuhan, China, which highlighted the need for improved prevention and control of TB in China. METHODS: Between 24 November and 9 December 2020 two milu deer and a northern pig-tailed macaque that were displaying signs of wasting died. Post-mortem, histopathological diagnosis and acid fast staining were used for the dead animals. Multiple PCR for Mycobacterium tuberculosis complex (MTBC) was performed to identify the bacterial in both milu deer and northern pig-tailed macaque. The serum antibody iELISA for MTBC was then performed for all the surviving milu deer and northern pig-tailed macaques. Six seropositive milu deer and a seropositive northern pig-tailed macaque were subsequently euthanised and, along with two other dead milu deer, necropsied. DNA from these tissue samples was extracted and detected MTBC using PCR and Real-time PCR. Subsequently bacterial isolation was used to confirm the infection. RESULTS: The lungs of the dead animals displayed gross and histological TB-like lesions and changes, and red staining bacilli were detected in smears of the lesions by microscopy after acid fast staining. Mycobacterium bovis (M. bovis) was detected in the two milu deer and Mycobacterium tuberculosis (M. tb) in the northern pig-tailed macaque using multiple PCR for MTBC. 35.3% surviving milu deer and 50% surviving northern pig-tailed macaques MTBC serologically positive. Six of the euthanised milu deer were also positive on a DNA test for M. bovis and the euthanised northern pig-tailed macaque was positive to M. tb. CONCLUSIONS: This is the first report of tuberculosis in the endangered species, milu deer and northern pig-tailed macaques, in China, and warrants urgent attention by researchers and conservation authorities. These cases highlight the need for expanding surveillance for MTBC to zoos in China.

Prevalence of Mycoplasma bovis Infection in Calves and Dairy Cows in Western Australia.

Mycoplasma bovis (M. bovis) can cause a multitude of diseases in cattle, with detrimental effects on the farm economy and the welfare of both adult and young cattle. The objective of this study was to determine the prevalence of M. bovis in adult cows and calves in the south-west region of Western Australia. A cross-sectional study was conducted on 29 dairy farms with 699 apparently healthy adult lactating cows and 495 young calves during 2019-2020. Nasal swabs and blood samples collected from the animals and bulk tank milk (BTM) samples were assessed for M. bovis-specific proteins and antibodies by using polymerase chain reaction (PCR) and Mycoplasma immunogenic lipase A- Enzyme-Linked Immune Sorbent Assay (MilA ELISA). A seroprevalence of 42.5% (95% CI: 38.9-46.2) and 61% (95% CI: 56.6-65.2) was found in adult lactating cows and calves, respectively. The herd-level seroprevalence of M. bovis ranged from 4% (95% CI: 07-19.5) to 92% (95% CI: 75.0-97.8) in adult lactating cows and 25% (95% CI: 10.2-49.5) to 87% (95% CI: 67.9-95.5) for calves in these farms. None of the BTM and nasal swab samples were positive for M. bovis, indicating an absence of any current active infections on the farms. The female calves and pure Holstein-Friesian animals are twice as likely to be seropositive for M. bovis compared to male calves (OR 2.4; 95% CI: 1.7-3.5) and Holstein-Friesian crossbred calves (OR 2.4; 95% CI: 1.7-3.5). The high seroprevalence in both adult and young cattle in the southwest dairy farms of Western Australia warrants more effective farm biosecurity measures and further evaluation of the current prevention and management measures practiced on the farms.

Knowledge, Attitudes, and Practices (KAPs) of Farmers on Foot and Mouth Disease in Cattle in Baghlan Province, Afghanistan: A Descriptive Study.

This study was performed to investigate the knowledge, attitudes, and practices (KAPs) of farmers, animal traders, and veterinary professionals on FMD in Baghlan province, Afghanistan. Four structured questionnaires were administered to the respondents. Almost half (48.5%) of the farmers had heard of the occurrence of FMD in their neighbourhood or knew the name of the disease. The majority of farmers could recognise the clinical signs of FMD in their animals (salivation, 85.9%; tongue ulcers, 78.8%; gum lesions, 78.2%; hoof lesions, 76.8%). Most farmers stated that the "introduction of new animals" was the primary cause of FMD appearing on their farms and to control the spread of the disease, over half of the farmers (56%) preferred not to buy cattle from unknown or potentially infected sources. Animal traders' knowledge was limited to recognising some clinical signs of the disease such as: salivation, and lesions in the mouth and on the feet. No animals were directly imported by the traders from outside Afghanistan. Over half of the local veterinary professionals (65%) kept record books of the animal diseases seen and/or treatment plans undertaken, and 80% of them reported the occurrence of FMD to the provincial, regional, and central veterinary authorities. No regular vaccination programme against FMD was implemented in the province. Poor import controls and quarantine were considered to be the main barriers to the control of FMD in the study area and the surrounding provinces. It can be concluded that, despite relatively good knowledge about FMD in the study area, there are gaps in farmers' and traders' knowledge that need to be addressed to overcome the burden of the disease in the province. These should focus on strengthening interprovincial quarantine measures and implementation of regular vaccination campaigns against the circulating FMDV within the area.

Seroprevalence and risk factors for foot-and-mouth disease in cattle in Baghlan Province, Afghanistan.

A serological study of 376 cattle from 198 herds and a concurrent survey of farmers were undertaken in 53 villages in Khinjan, Doshi and Puli Khumri districts of Baghlan province, Afghanistan to determine the seroprevalence of Foot and Mouth Disease (FMD) and to identify risk factors for seropositive herds. A total of 419 cases of FMD were reported by the farmers in the year preceding the survey. The animal-level population seroprevalence was estimated at 42.0% (95% CI, 37.0-47.2). The seroprevalence increased with age in the sampled cattle (<2 years - 30.4%, 2-6 years - 40.3% and >6 years - 52.2%). Herds were more likely to be seropositive if the farmers: had purchased cattle in the year prior to the survey (OR = 2.6; 95% CI, 1.37-4.97); purchased ruminants from unknown (potentially risky) sources (OR = 2.13; 95% CI, 1.13-4.03); and sold milk to the market (OR = 1.99; 95% CI, 1.09-3.63). Herds that had been vaccinated had a lower odds of being seropositive (OR = 0.33; 95% CI, 0.68-0.66). This was the first epidemiological study of FMD in Baghlan province and the findings provide valuable direction for disease control on FMD in this and other provinces in Afghanistan.

History and epidemiology of foot-and-mouth disease in Afghanistan: a retrospective study.

BACKGROUND: Foot and mouth disease (FMD) is endemic in Afghanistan with serotypes O, A and Asia 1 being prevalent. A retrospective study of data collected through passive surveillance of outbreaks of FMD in Afghanistan from 1995 to 2016 was undertaken to determine the temporal and spatial distribution of FMD in the country. RESULTS: A total of 4171 outbreaks were reported between 1995 and 2008 with a strong correlation between the number of outbreaks and the number of provinces (r = 0.85, s = 68.2, p < 0.001); and between the number of outbreaks and the number of districts containing infected animals (r = 0.68, s = 147.8, p = 0.008). Of 7558 samples collected from livestock originating from 34 provinces in 2009, 2011 and 2013-2015, 54.1% were test positive (FMDV 3ABC-trapping ELISA) and the prevalence varied significantly between years (χ2 = 263.98, df = 4, P < 0.001). Clinically suspected cases were reported in 2016 with a substantial positive correlation (r = 0.70, P < 0.001) between the number of districts with cases and the number of reported cases. Serotype O was the predominant serotype detected during the study period, although serotypes A and Asia1 were also detected. Cattle were involved in all outbreaks in the study period and infections were detected in all years of the study in Hirat province in the north-west (bordering Iran), Nangarhar province in the east (bordering Pakistan) and Kabul province in the centre of the country. CONCLUSIONS: The current paper was the first analysis of existing data focusing on the spatiotemporal distribution of FMD in Afghanistan. The findings from this study provide valuable direction for further research to understand the epidemiology of FMD and its control in Afghanistan.

Knowledge, attitudes and practices (KAP) towards rabies and free-roaming dogs (FRD) in Shirsuphal village in western India: A community based cross-sectional study.

The lack of awareness about dog-bite related rabies in the rural population of developing countries, including India, is a major impediment to controlling the incidence of disease in humans. A survey of 127 rural residents was undertaken in Shirsuphal village in western India using a structured questionnaire to assess the influence of demographic and pet/livestock owning characteristics on the knowledge, attitudes and practices of the respondents towards rabies and free roaming dogs (FRD). Multivariable logistic regression models were constructed and the knowledge of the rural residents of Shirsuphal village was found to be significantly influenced by family size (OR 2.1, 95%CI 1.0-4.6, p = 0.04) and poultry ownership (OR 2.3, 95%CI 1.1-4.9, p = 0.03), while their attitudes towards FRD was significantly influenced by age of the respondents (OR 2.6, 95% CI 1.2-5.8) and ownership of cattle/buffalo (OR 2.2, 95% CI 1.1-5.5). Although the knowledge score about rabies was high, a comprehensive understanding of the disease was lacking. Concerted efforts to widen the knowledge about rabies and promote healthier practices towards FRD are recommended.

Knowledge, attitudes and practices towards dog-bite related rabies in para-medical staff at rural primary health centres in Baramati, western India.

The lack of awareness regarding rabies amongst rural primary care health staff and their adverse practices towards the management of dog-bite wounds is a major contributor to the high incidence of rabies infection and subsequent human mortality in India. A Knowledge, Attitudes and Practices survey was carried out involving 54 nursing and non-nursing staff working in 18 rural Primary Health centres and sub-centres around Baramati town of Pune district in Western India. Multivariable logistic regression models were constructed to assess factors that influenced knowledge of rabies and practices towards management of dog-bite related wounds. The more experienced and better-educated workers were found to have a good awareness of rabies (OR 3.4, 95%CI 1.0-12.1) and good practices towards dog-bite wound management (OR 5.6, 95%CI 1.2-27.0). Surprisingly, non-nursing staff were significantly more knowledgeable about rabies (OR 3.5, 95%CI 1.0-12.3), but their practices towards dog-bite wound management were inadequate (OR 0.18, 95%CI 0.04-0.8) compared to the nursing staff. It is recommended that a mandatory training module for primary care health staff be developed and implemented to improve their knowledge regarding rabies and management of dog-bite wounds to reduce the incidence of human rabies in rural India.

A Comparative Study of Enumeration Techniques for Free-Roaming Dogs in Rural Baramati, District Pune, India.

The presence of unvaccinated free-roaming dogs (FRD) amidst human settlements is a major contributor to the high incidence of rabies in countries such as India, where the disease is endemic. Estimating FRD population size is crucial to the planning and evaluation of interventions, such as mass immunisation against rabies. Enumeration techniques for FRD are resource intensive and can vary from simple direct counts to statistically complex capture-recapture techniques primarily developed for ecological studies. In this study we compared eight capture-recapture enumeration methods (Lincoln-Petersen's index, Chapman's correction estimate, Beck's method, Schumacher-Eschmeyer method, Regression method, Mark-resight logit normal method, Huggin's closed capture models and Application SuperDuplicates on-line tool) using direct count data collected from Shirsuphal village of Baramati town in Western India, to recommend a method which yields a reasonably accurate count to use for effective vaccination coverage against rabies with minimal resource inputs. A total of 263 unique dogs were sighted at least once over 6 observation occasions with no new dogs sighted on the 7th occasion. Besides this direct count, the methods that do not account for individual heterogeneity yielded population estimates in the range of 248-270, which likely underestimate the real FRD population size. Higher estimates were obtained using the Huggin's Mh-Jackknife (437 ± 33), Huggin's Mth-Chao (391 ± 26), Huggin's Mh-Chao (385 ± 30), models and Application "SuperDuplicates" tool (392 ± 20) and were considered more robust. When the sampling effort was reduced to only two surveys, the Application SuperDuplicates online tool gave the closest estimate of 349 ± 36, which is 74% of the estimated highest population of free-roaming dogs in Shirsuphal village. This method may thus be considered the most reliable method for estimating the FRD population with minimal inputs (two surveys conducted on consecutive days).

Knowledge, Attitudes, and Practices of 50 Patients with Surgically Treated Cystic Echinococcosis from Basrah Province, Iraq.

Cystic echinococcosis (CE) is recognised worldwide as a neglected disease of public health concern, particularly in low- and middle-income countries. The objectives of this study were to describe, using a questionnaire survey, the characteristics, attitudes, knowledge, and practices of patients with CE from Basrah Province, Iraq. We interviewed 50 patients, 31 female and 19 male, of whom, 74% originated from rural areas. Approximately half (48%) of the participants reported slaughtering livestock at home for their families' consumption; 78% indicated the presence of a large number of stray dogs roaming freely about their village; 86% reported that they never boiled water prior to drinking it; and 26% reported not washing vegetables prior to eating them. Although a large proportion of the participants (72%) had heard of hydatid disease prior to becoming ill, over half (57%) were not aware how the disease was transmitted from animals to humans. This study highlighted a gap in health education efforts regarding CE in Southern Iraq, with a lack of counselling of patients on how to prevent reinfection. An intensive control programme should be implemented and founded on health education to reduce CE disease in Basrah.

Canine vector-borne infections in Mauritius.

BACKGROUND: Canine vector-borne diseases have a worldwide distribution, but to the best of our knowledge, no research has been carried out to evaluate their presence on the Indian Ocean island of Mauritius. An investigation into canine vector-borne infections was conducted in dogs (n = 78) resident at an animal shelter in Port Louis, Mauritius using a combination of traditional microscopy and serological methods. METHODS: Ticks were manually collected from the stray dog population for identification as well as for quantifying tick burdens. Blood was also collected from each dog via either the jugular vein or the cephalic vein, and was stored in EDTA tubes. The stored blood was then used to measure PCV values, make blood smears for the identification of parasites, and used for serological testing of vector-borne disease. RESULTS: A total of 178 ticks were collected from 52 dogs and identified as Rhipicephalus sanguineus (175/178) or Amblyomma variegatum (3/178). Twenty-six (33%; 95% CI 23, 45) dogs were seropositive for Ehrlichia spp., and 12 (15%; 95% CI 8, 25) for Anaplasma spp., Dirofilaria antigen was detected in 14 (18%; 95% CI 10, 28), and nine (12%; 95% CI 5, 21) dogs had Hepatozoon canis gamonts observed in blood films during microscopic examination. Eleven (14%; 95% CI 7, 24) dogs were co-infected with two pathogens. Borrelia burgdorferi antibodies were not detected in any dogs. CONCLUSIONS: Infection with these pathogens had no significant effect on the packed cell volume (PCV), but high tick burdens were significantly associated with the presence of a tick-borne pathogen. This is the first study of its kind on the dog population in Mauritius and demonstrates the presence of previously undocumented canine vector-borne infections on the island. The relatively high proportion of infected dogs within the study should alert clinicians to the presence of canine vector-borne diseases on the island of Mauritius.

Outbreaks of foot-and-mouth disease in Peninsular Malaysia from 2001 to 2007.

This is a retrospective study of the outbreaks of foot-and-mouth disease (FMD) in Peninsular Malaysia between 2001 and May 2007. In total, 270 outbreaks of FMD were recorded. Serotype O virus (89.95 %) and serotype A (7.7 %) had caused the outbreaks. Significant differences on the occurrence of FMD were found between the years (t = 5.73, P = 0.000, df = 11), months (t = 4.7, P = 0.000, df = 11), monsoon season (t = 2.63, P = 0.025, df = 10) and states (t = 4.84, P = 0.001, df = 10). A peak of outbreaks observed in 2003 could be due to increased animal movement and the other peak in 2006 could be due to a compromised FMD control activities due to activities on the eradication of highly pathogenic avian influenza. Cattle (86 % of outbreaks) suffered the most. However, no difference in disease occurrence between species was observed. The populations of cattle (r = 0.672, P = 0.023) and sheep (r = 0.678, P = 0.022) were significantly correlated with occurrence of FMD. Movement of animals (66 % of outbreaks) was the main source for outbreaks. A combination of control measures were implemented during outbreaks. In conclusion, the findings of this study show that FMD is endemic in Peninsular Malaysia, and information gained could be used to improve the existing control strategy.

A retrospective study on the epidemiology of foot-and-mouth disease in Bhutan.

A retrospective study on the outbreaks of foot-and-mouth disease (FMD) in Bhutan, between the years 1996 and 2008, based on the data collected through passive surveillance, was undertaken. A total of 230 outbreaks of FMD at sub-district level were recorded in 299 villages located in 19 out of the 20 districts in the country. There were no significant differences between the years (P = 0.998) or months (P = 0.989) on the incidence of FMD. The sub-districts in the north (altitude >1,000 m above mean sea level) had significantly (P = 0.008) higher incidences of outbreaks in winter than in summer. The sub-districts that shared border with India had significantly more outbreaks than those that didn't (P = 0.001). Cattle were the most predominant species affected being involved in all of the outbreaks reported. Serotype O, which constituted 70.6% of the outbreaks typed was the most predominant serotype prevalent in Bhutan followed by A (16.7%), Asia 1 (8.8%), and C (3.9%). Cattle density was significantly positively correlated (P = 0.023) with the incidence of disease. Three waves of outbreaks of epidemic proportions were reported in 1997/1998, 2002/2003, and 2007/2008 due to the PanAsia strain of the O serotype. The study highlights the incursion of the PanAsia strain of the O serotype into the country, possibly, through the transboundary movement of animals and the need for active surveillance of FMD, especially at the border areas. The study also highlights the significance of the O serotype and cattle as the main indicator species in the epidemiology of FMD in Bhutan. The findings from this study can be used as baseline epidemiological data for further research to understand the epidemiology of FMD in Bhutan.