Spatial and temporal analysis of haemorrhagic septicaemia outbreaks in India over three decades (1987-2016).

Haemorrhagic septicaemia (HS) is an economically important disease affecting cattle and buffaloes and the livelihoods of small-holder farmers that depend upon them. The disease is caused by Gram-negative bacterium, Pasteurella multocida, and is considered to be endemic in many states of India with more than 25,000 outbreaks in the past three decades. Currently, there is no national policy for control of HS in India. In this study, we analysed thirty year (1987-2016) monthly data on HS outbreaks using different statistical and mathematical methods to identify spatial variability and temporal patterns (seasonality, periodicity). There was zonal variation in the trend and seasonality of HS outbreaks. Overall, South zone reported maximum proportion of the outbreaks (70.2%), followed by East zone (7.2%), Central zone (6.4%), North zone (5.6%), West zone (5.5%) and North-East zone (4.9%). Annual state level analysis indicated that the reporting of HS outbreaks started at different years independently and there was no apparent transmission between the states. The results of the current study are useful for the policy makers to design national control programme on HS in India and implement state specific strategies. Further, our study and strategies could aid in implementation of similar approaches in HS endemic tropical countries around the world.

Viral Haemorrhagic Septicemia Virus (VHSV) Isolated from Atlantic Herring, Clupea harengus, Causes Mortality in Bath Challenge on Juvenile Herring.

Viral hemorrhagic septicaemia virus (VHSV) has been demonstrated to cause high mortalities in a wide range of teleosts, farmed as well as wild. In Europe, VHSV of genotypes Ib, Id, II, and III have been detected in wild fish, including Atlantic herring Clupea harengus, but disease outbreaks have not been observed in Atlantic herring and the effects on wild stocks are not well documented. Here, we have tested two VHSV isolates from herring (genotypes Ib and III, from the western coasts of Norway and Denmark, respectively) in a challenge experiment with herring (mean weight 2.59 g, SD 0.71 g) caught on the west coast of Denmark. The Norwegian genotype Ib isolate (NO-F-CH/2009) showed an accumulated mortality of 47% compared to 6% mortality with the Danish genotype III isolate 4p168 and zero in the unchallenged control group. In both groups, we found positive rt-RT-PCR and positive immunohistochemistry of VHSV from days 6 and 8 onward. With both isolates, the organs mainly affected were the heart and kidney. The results demonstrate the susceptibility of Atlantic herring to VHSV, and both genotypes gave pathological findings in several organs. Genotype III showed a low mortality rate, and the importance of this genotype for herring is therefore not determined. Genotype Ib showed both high prevalence and mortality, and this genotype is therefore likely to have a negative effect on wild Atlantic herring stocks. Further examinations to determine how VHSV can affect wild Atlantic herring stocks are needed.

First outbreak of bovine haemorrhagic septicaemia caused by Pasteurella multocida type B in Spain - Short communication.

This paper describes the first documented outbreak of haemorrhagic septicaemia (HS) caused by Pasteurella multocida type B in cattle in Spain. This acute, highly fatal septicaemia causes major economic losses in cattle and buffaloes in many areas of Asia and Africa. In other species and in European countries it is an infrequently reported disease. Acute septicaemic pasteurellosis occurred in a free-range farm of 150 cattle and 70 beef calves in Southern Spain. Twenty-one calves and one cow were affected, of which three calves and the adult cow died. Postmortem examination revealed characteristic oedema in the ventral area of the neck and the brisket region, and widespread haemorrhages in all organs. Pure cultures of P. multocida were obtained from all tissues and organs studied. The aetiological agent was further confirmed by molecular and biochemical analysis as P. multocida capsular type B, biovar 3. Although the source of infection could not be determined, wildlife may play an important role. The use of tulathromycin in the initial stage of the disease might be related to the low morbidity and mortality of this outbreak. After using an autogenous vaccine no more cases of HS were observed.

Improving Village Animal Health Worker participation in national disease surveillance systems: A case study from Cambodia.

Para-veterinary systems have arisen in numerous developing countries to address the low capacity of national veterinary services in meeting livestock health demands of mainly smallholder farmers. In Cambodia, the village animal health worker (VAHW) system was established in the early 1990s, involving short training programmes to equip VAHWs to provide basic animal health services for smallholder farmers, particularly the vaccination of cattle for haemorrhagic septicaemia (HS). However, there are increasing expectations that VAHWs provide village level disease surveillance information to the national veterinary services, despite their low-level disease diagnostic skills. To identify opportunities to improve the disease reporting system in Cambodia, a closed-ended cross-sectional study of VAHWs (n = 80) from two provinces was conducted in 2015, examining their contact frequency with district and provincial animal health authorities. Ordinal logistic regression was used to determine factors associated with increased frequency of VAHW contact with the district animal health authorities responsible for national disease reporting. Positive associations between income generated from VAHW activities (p = .01) and the frequency of visiting farmers (p < .0001), with more frequent contacts with district animal health authorities, were identified. High levels of inappropriate use of antibiotics, with almost 90% of VAHW reporting they use antibiotics to treat FMD-affected animals, were reported. Although further research to increase the rates of reporting at the smallholder farmer and VAHW levels is required, increased emphasis on complying with the World Organisation for Animal Health's competency guidelines for veterinary paraprofessionals may be required if they are expected to contribute to national animal disease surveillance systems.

Isolation characterization, virulence potential of Weissella ceti responsible for weissellosis outbreak in rainbow trout (Oncorhynchus mykiss) cultured in Mexico.

Weissella ceti, a Gram-positive nonmotile bacterium, is currently an emerging pathogen within rainbow trout (Oncorhynchus mykiss) farms in China, Brazil, the United States, and Japan. This study is the first to isolate, identify, and characterize W. ceti isolates from rainbow trout farmed in Mexico. In late 2015, a severe disease outbreak caused a 60% mortality rate among 20,000 fish. The diseased rainbow trout (100-300 g average) exhibited severe cachexia, body darkening, abdominal distension, exophthalmia, haemorrhages, and corneal opacity. Internally, diseased fish had pale gills; multifocal, disseminated whitish spots on the liver; haemorrhages in the swim bladder, ovary, and on the parietal surface of the muscle; and hearts with pseudo-membrane formation. Histologically, lesions were characterized by corneal oedema, degenerative and necrotic hepatitis, and meningitis. A brain (W-1) and kidney (W-2) isolate were identified as W. ceti through polyphasic taxonomy, which included phenotypic characterization and 16S rRNA sequencing. RAPD and ERIC-PCR analyses demonstrated genetic homogeneity among the Mexican isolates. Virulence tests in rainbow trout through intraperitoneal W. ceti injections at concentrations of 1 × 104 , 1 × 105 , and 1 × 106 CFU per fish resulted in cumulative mortality rates of 25%, 62.5%, and 87.5%, respectively, as well as the same clinical signs of hemorrhagic septicaemia as were recorded for the natural outbreak. The present report is the first to confirm the presence of W. ceti in Mexico, thus extending the known geographical distribution of this pathogen across the Americas.

SURVEILLANCE FOR HEMORRHAGIC SEPTICEMIA IN BUFFALO ( BUBALUS BUBALIS) AS AN AID TO RANGE EXPANSION OF THE JAVAN RHINOCEROS ( RHINOCEROS SONDAICUS) IN UJUNG KULON NATIONAL PARK, INDONESIA.

The Javan rhinoceros ( Rhinoceros sondaicus) of Ujung Kulon National Park (UKNP) is the crown jewel of Indonesia's rich natural history. The park lies on a peninsula surrounded by coastline and agriculture-dominated landscapes. The invasion of water buffalo ( Bubalus bubalis) into the park carries a substantial health risk to the Javan rhinoceros and threatens plans to establish a new population outside of its only current range in UKNP. Hemorrhagic septicemia (HS), known locally as septicemia epizootica and caused by Pasteurella multocida B:2, could thwart Indonesia's efforts to expand the range of the Javan rhinoceros. Because HS was considered eradicated from Banten Province, few preventative programs have been available to farmers. During June 2012-July 2013, biologic samples were collected from 770 water buffalo in 19 villages. Deep nasal swabs ( n=85) were taken for bacterial culture and blood samples ( n=770) were collected for serologic testing. No animals were positive on culture. The prevalence of antibody to P. multocida in this population was 1.8% (14 of 770 animals). A structured questionnaire was used to gather information about possible risk factors. Husbandry practices associated with presence of antibody in water buffalo included lack of a permanent area to house buffalo at night, low body condition score (=2), high body temperature (≥40 C), a history of clinical signs or sudden death in the previous year, and a grazing system that utilized significant forage inside the park. Antibody was not associated with sex, age, vaccination status, or season. Understanding HS disease dynamics in the buffalo adjacent to UKNP may improve the livelihoods of people and health of endangered rhinoceroses in this ecosystem.

Epidemiological features and financial loss due to clinically diagnosed Haemorrhagic Septicemia in bovines in Karnataka, India.

The epidemiological features and financial losses due to Haemorrhagic Septicemia (HS) in bovines were studied in Karnataka state using the primary data collected from 133 clinically diagnosed HS affected farms. The various losses due to HS and the Benefit- Cost of the vaccination programme in cattle and water buffaloes were studied using mathematical models. The number of HS outbreaks were higher during the year 2002 and peaked during 2005 and thereafter declined due to targeted vaccination against HS. The morbidity and mortality risks were lower in large farms than medium and small farms, and lower in indigenous cattle compared to high yielding crossbred cattle and water buffaloes. The disease occurrence was more in in-milk animals causing serious economic loss to the farmers. Most outbreaks were observed during monsoon season, though the disease was prevalent throughout the year. The mean milk loss per animal was $2, $11 and $50 in indigenous cattle, water buffaloes and crossbred cattle, respectively. In the case of draught animals, the average effective draught power was unavailable for 1.2days/outbreak resulting in a loss of $5 per affected oxen. The treatment and extra labor expenses incurred per animal were $24 and $7, respectively. The average loss per animal due to mortality loss was $275, $284 and $415 in case of indigenous cattle, water buffaloes and crossbred cattle, respectively. The projected loss for the state of Karnataka were $23.89, $17.92 and $11.95 million under high, medium and low HS incidence scenarios, respectively. The Benefit Cost Analysis (BCA) of the vaccination against HS has been estimated at 5.97:1, 4.48:1 and 2.98:1 under high, medium and low incidence scenarios, respectively. The results highlight the important epidemiological features and financial losses to the affected households and the state of Karnataka.

Re-emergence of bovine haemorrhagic septicaemia in Hungary.

This paper reports an outbreak of haemorrhagic septicaemia caused by Pasteurella multocida B:2 in beef calves, a disease that has not been described in the Hungarian literature since 1943, and has not been reported to the World Organisation For Animal Health (OIE) since 1970. Acute haemorrhagic septicaemia was confirmed in beef calves on one small farm, and was suspected on two further nearby holdings with concomitant unexplained losses. The source of the infection could not be determined. Apart from a short duration of depression and loss of appetite, the affected calves developed characteristic distal limb oedema. Gross findings in two calves submitted for laboratory examinations included subcutaneous oedema and haemorrhages on serous membranes, and in one case severe pharyngeal lymph node enlargement was observed. Histological examinations revealed lesions characteristic of septicaemia. Moderate to large amounts of Pasteurella antigens were detected in all organs tested by immunohistochemistry. Two isolates of P. multocida (Pm240, Pm241) were cultured from these cases and examined in detail. These were identified as P. multocida ssp. multocida biovar 3. Both were toxA negative and belonged to serotype B:2. Multilocus sequence typing was used to assign these to a new sequence type (ST64) that is closely related to other haemorrhagic septicaemia causing strains of P. multocida regardless of the host.

A Case-control Study of Haemorrhagic Septicaemia in Buffaloes and Cattle in Karachi, Pakistan, in 2012.

A retrospective epidemiological case-control study was performed in Karachi, Pakistan, from January to April 2013. The owners of 217 dairy cattle and buffalo farms from six different locations in Karachi were interviewed. The aim of the study was to identify risk factors associated with the presence of haemorrhagic septicaemia (HS). Farms with a history of at least one instance of sudden death in a dairy animal during 2012 and a positive clinical HS diagnosis (made by local veterinarians) were defined as cases. Farms having no history of sudden deaths in 2012 were defined as controls. Univariable analyses were initially conducted, and factors with P ≤ 0.25 were offered to a multivariable logistic regression model to identify putative risk factors. The final multivariable logistic model contained five factors. Vaccination was found to be a protective factor (OR = 0.22) along with the length of time cattle were kept on farm (months). For every extra month cattle were kept, the odds of HS disease were reduced by a factor of 0.9. In contrast, for every extra animal in a herd, the risk of infection increased by a factor of 1.01. Supplying underground water and the presence of foot and mouth disease on the farm increased the risk by 2.90 and 2.37, respectively. To understand the epidemiology of HS in Karachi dairy herds, more in-depth research is required to study the risk and protective factors identified in this survey and to evaluate risk mitigation strategies, where possible.

Molecular Epidemiology of Pasteurella multocida Circulating in India by Multilocus Sequence Typing.

Multilocus sequence typing (MLST), a sequence-based typing method for bacterial pathogens, is currently the best method for long-term epidemiological study and to understand the population structure of the bacteria. This investigation was carried out to study the diversity of Pasteurella multocida isolates circulating in India. Ten different sequence types (ST) identified in this study are ST 122 from cattle, goat, mithun and pig; ST 50 from pig; ST 9 from cattle and sheep; ST 229 from cattle and goat; ST 71 and ST 277 from cattle; and ST 129, ST 280, ST 281 and ST 282 from avian species. Of these, ST 277, ST 280, ST 281 and ST 282 were identified for the first time. The analysis of results provides novel epidemiological information on the circulation of multiple STs across India. The majority of STs or their variants identified in this study have already been reported from different parts of the globe. This suggests that probably transboundary spread of strains across countries and continents has occurred across evolutionary time and is still happening. The isolation of ST 122 from small ruminants and pigs suggests that these species may be included in the preventive vaccination policy for effective control of haemorrhagic septicaemia in India.

Serotyping of foot and mouth disease virus and Pasteurella multocida from Indian gaurs (Bos gaurus), concurrently infected with foot and mouth disease and haemorrhagic septicaemia.

We report the serotyping of foot-and-mouth disease virus (FMDV) and Pasteurella multocida from Indian gaurs which were concurrently infected with foot-and-mouth disease (FMD) and haemorrhagic septicaemia. Bannerghatta biological park (BBP), a national park located in the outskirts of Bengaluru city, Karnataka, India, is bordered by several villages. These villages witnessed massive outbreaks of FMD which spread rapidly to the herbivores at BBP. Post-mortem was conducted on carcasses of two Indian gaurs that died with symptoms of FMD. The salient gross findings included extensive vesicular lesions on the tongue, gums, cheeks, upper palate and hooves. Haemorrhagic tracheitis and ecchymotic haemorrhages on the heart were characteristic. The vesicular lesions of oral cavity were positive for 'O' type of FMD virus by sandwich enzyme-linked immuno sorbent assay (ELISA). The heart blood and spleen samples yielded growth of pure cultures of P. multocida. The isolates were typed as P. multocida type B using KTSP61 and KTT72 primers yielding specific amplicons of 620 bp. The phylogenetic analysis of the isolates was carried by sequencing of 1.4-Kbp nucleotides on the 16S ribosomal RNA (rRNA) gene of the isolates.

The Socioeconomic Impacts of Clinically Diagnosed Haemorrhagic Septicaemia on Smallholder Large Ruminant Farmers in Cambodia.

Haemorrhagic septicaemia (HS) is an acute fatal infectious disease of mainly cattle and buffalo and outbreaks occur commonly in Cambodia. Disease outbreak reports were examined to select five villages from three provinces for a retrospective investigation of HS epidemiology and socioeconomic impact on smallholders, with an aim of identifying potential benefits from improving disease prevention through biosecurity and vaccination. The Village Animal Health Worker (VAHW) or Chief in each village and 66 affected smallholders were surveyed. At the village level, 24% of all households were affected with an estimated mean village herd morbidity of 10.1% and mortality of 28.8%. Affected farmers reported HS disease morbidity and mortality at 42.7% and 63.6% respectively. Buffalo had a higher morbidity (OR = 2.3; P = 0.003) and mortality (OR = 6.9; P < 0.001) compared with cattle, and unvaccinated large ruminants a higher morbidity (OR = 2.9; P = 0.001). The financial impact varied depending on whether the animal survived, provision of treatment, draught replacement and lost secondary income. The mean cost per affected household was USD 952.50 based on ownership of five large ruminants. The impact per affected animal was USD 375.00, reducing the pre-disease value by 66.1%. A partial budget revealed an overwhelming incentive for farmers to practice biannual vaccination, with a net benefit of USD 951.58 per household based on an annual disease incidence rate of 1. Sensitivity analysis showed that a net benefit of USD 32.42 remained based on an outbreak every 20 years. This study indicates HS can cause a catastrophic financial shock to smallholders and remains a critical constraint to improving large ruminant productivity and profitability. Addressing HS disease control requires a focus on improving smallholder farmer knowledge of biosecurity and vaccination and should be priority to stakeholders interested in addressing regional food insecurity and poverty reduction.

A retrospective study on the epidemiology of anthrax, foot and mouth disease, haemorrhagic septicaemia, peste des petits ruminants and rabies in Bangladesh, 2010-2012.

Anthrax, foot and mouth disease (FMD), haemorrhagic septicaemia (HS), peste des petits ruminants (PPR) and rabies are considered to be endemic in Bangladesh. This retrospective study was conducted to understand the geographic and seasonal distribution of these major infectious diseases in livestock based on data collected through passive surveillance from 1 January 2010 to 31 December 2012. Data analysis for this period revealed 5,937 cases of anthrax, 300,333 of FMD, 13,436 of HS, 247,783 of PPR and 14,085 cases of dog bite/rabies. While diseases were reported in almost every district of the country, the highest frequency of occurrence corresponded to the susceptible livestock population in the respective districts. There was no significant difference in the disease occurrences between districts bordering India/Myanmar and non-border districts (p>0.05). Significantly higher (p<0.01) numbers of anthrax (84.5%), FMD (88.3%), HS (84.9%) and dog bite/rabies (64.3%) cases were reported in cattle than any other species. PPR cases were reported mostly (94.8%) in goats with only isolated cases (5.2%) in sheep. The diseases occur throughout the year with peak numbers reported during June through September and lowest during December through April, with significant differences (p<0.01) between the months. The annual usages of vaccines for anthrax, FMD, HS and PPR were only 7.31%, 0.61%, 0.84% and 11.59% of the susceptible livestock population, respectively. Prophylactic vaccination against rabies was 21.16% of cases. There were significant differences (p<0.01) in the administration of anthrax, FMD and HS vaccines between border and non-border districts, but not PPR or rabies vaccines. We recommend that surveillance and reporting of these diseases need to be improved throughout the country. Furthermore, all suspected clinical cases should be confirmed by laboratory examination. The findings of this study can be used in the formulation of more effective disease management and control strategies, including appropriate vaccination policies in Bangladesh.

Molecular typing of haemorrhagic septicaemia-associated Pasteurella multocida isolates from Pakistan and Thailand using multilocus sequence typing and pulsed-field gel electrophoresis.

A comparative genetic study of 23 field isolates and vaccine strains of Pasteurella multocida associated with haemorrhagic septicaemia cases from Pakistan and Thailand was done using pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The MLST sequence type (ST) for all 20 of the 23 isolates tested was 122. The PFGE results showed one band difference between the Pakistani and the Thai isolates. Sequence type 122 is the dominant associated profile with haemorrhagic septicaemia (HS) cases in South Asia. The study supports the concept of using PFGE for short-term epidemiology and MLST for long-term epidemiology.

[Acute pasteurellosis in fallow deer, cattle and pigs in a region of Eastern Germany]. / Akute Pasteurellose bei Damwild, Rindern und Schweinen in einer Region im Osten Deutschlands.

Hemorrhagic septicaemia, an acute disease caused by P multocida capsular type B which is rarely detected in Europe, caused considerable losses in fallow deer, cattle and pigs within a region along the border of the federal states Brandenburg and Saxony-Anhalt in the summer of 2010. Clinical appearances and diagnostic findings are presented and possible triggering influences discussed. Pasteurella multocida capsular type B has not been cultivated from clinically healthy cattle and pigs of the region. Examination of fallow deer and roe deer in the region revealed the presence of singular carriers, which may act as a source of clinical infections.

Weissella sp. outbreaks in commercial rainbow trout (Oncorhynchus mykiss) farms in Brazil.

The genus Weissella contains 14 bacterial species that usually occur in nutrient-rich environments and in fermented foods and beverages. Outbreaks of hemorrhagic septicemia were reported in three commercial rainbow trout (Oncorhynchus mykiss) farms in Brazil in 2008 and 2009. Seventy-seven Gram-positive isolates were obtained from 41 diseased fish from these farms. The bacterial strains were identified as Weissella at the genus level using biochemical tests, Weissella genus-specific PCR, and 16S rRNA sequencing. To evaluate potential routes of infection, rainbow trout juveniles were experimentally infected with the pathogen. In addition, the resistance of the pathogen to five antibiotics was tested, and provisional epidemiological cut-off values were calculated using the normalized resistance interpretation (NRI). All isolates presented similar phenotypic profiles and positive reactions for Weissella genus-specific PCR. The 16S rRNA sequences of the Brazilian strains showed 100% similarity with sequences of Chinese isolates that previously were identified as the first case of Weissella sp. infection in fish. The disease was successfully reproduced in the laboratory by intraperitoneal injection, immersion, and cohabitation between diseased and healthy fish. All isolates were resistant to sulfonamide, and based on NRI analysis, one, two, and three isolates were classified as non-wild-type (NWT) for erythromycin, oxytetracycline, and norfloxacin, respectively. This is the first description of multiple cases of Weissella sp. infection in rainbow trout farms outside of China, of infectious routes for the disease, and of provisional epidemiological cut-off values for resistance of these bacteria to four antibiotics.

The socio-economic impact of important camel diseases as perceived by a pastoralist community in Kenya.

This paper presents the results of a study conducted in a pastoral community in Kenya using participatory appraisal approaches. The objective of the study was to assess the socio-economic impact of camel trypanosomosis (surra) according to the perceptions of the pastoralists. Four livestock grazing units were conveniently selected and in each of them, three groups of key informants comprising five to eight persons were selected for the participatory exercises. Five camel diseases were listed in order of importance according to their severity and frequency of occurrence including trypanosomosis, mange, non-specific diarrhoea, tick infestations and haemorrhagic septicaemia. The losses listed as incurred due to the five diseases were: losses in milk, meat, blood, fats and hides, dowry payments, and depreciation in sale of animals, losses due to infertility and abortions, and losses due to the cost of treatment. There was good agreement (P < 0.05) between the informant groups on the losses incurred as a result of the diseases for all the selected loss indicators. Surra and mange were given high median scores on all the indicators while non-specific diarrhoea, tick infestations, and haemorrhagic septicaemia received moderate median scores. Based on the study findings it is concluded that the camel plays a central role in the lives of Turkana pastoralists and that surra has a devastating social and economic impact. There is a need for veterinary and policy decision-makers to focus more attention on the control of surra in this arid and semi-arid area of Kenya.

Community perceptions of important camel diseases in Lapur Division of Turkana District, Kenya.

This paper presents the results of a study conducted in Lapur Division of Turkana District, Kenya, to estimate the incidence and mortality of camel trypanosomosis using participatory methods. Four livestock camps ('adakars') were conveniently selected for the study. Four informant groups comprising 6 8 key persons were used for the participatory exercises. The camel diseases identified by the pastoralists in their order of importance according to annual incidence were: trypanosomosis (11.4%); mange (10.8%); tick infestation (7.9%); haemorrhagic septicaemia (7.7%); and non-specific diarrhoea (7.6%). Almost half (49.3%) of the camel population suffered from at least one disease over the previous year. The annual incidence and mortality rates of trypanosomosis were estimated at 15%, and 9.9%, in adult camels and 6.9% and 5.2%, in young camels, respectively. There was a seasonal occurrence of trypanosomosis, with most cases reported in the dry season. The prevalence levels of the disease reportedly declined from about 100%, in 1978 to an almost stable state of about 15% in 2002. This study revealed that camel trypanosomosis is still an important disease in Turkana District, exacting a heavy toll in terms of morbidity and mortality. The economic losses due to the disease were likely to have been great owing to the central role the camel plays in this arid district of Kenya.