Hepatitis E virus infection of slaughtered healthy pigs in Brazil

Hepatitis E virus (HEV) is an emerging pathogen that can be transmitted through contaminated raw or undercooked meat derived from domestic pigs. HEV infections have been documented among pig herds, pig products and environmental samples raising concern about the spread of the virus. HEV genotypes 3 and 4 are considered zoonotic and have been linked to human cases. HEV was detected in 51 of 335 bile samples (15.2%) from healthy pigs in Minas Gerais, Brazil. Phylogenetic analysis of partial sequences from ORF1 and ORF2 regions yielded discordant results, assigning isolates to subtypes 3c and 3i, respectively, suggesting intragenotypic HEV recombination.     

Hepatitis E virus infection in swine workers: A meta‐analysis

Hepatitis E virus (HEV) infects both humans and animals. Swine has been confirmed to be the principal natural reservoir, which raises a concern that HEV infection would be substantially increasing among swine workers. The present study calculated the pooled prevalence of IgG antibodies against HEV among swine workers and the general population in previous cross‐sectional studies. We conducted a meta‐analysis comparing the prevalence of HEV infection between swine workers and the general population, including local residents, blood donors and non‐swine workers. Through searches in three databases (PubMed and OVID in English, and CNKI in Chinese) and after study selection, a total of 32 studies from 16 countries (from 1999 through 2018) were included in the meta‐analysis. A random‐effect model was employed in the study; an I ² statistic assessed heterogeneity, and the Egger's test detected publication bias. The comparative prevalence of anti‐HEV IgG was pooled from the studies. Compared to the general population, the prevalence ratio (PR) for swine workers was estimated to be 1.52 (95% CI 1.38–1.76) with the I ² being 71%. No publication bias was detected (p = 0.40). A subgroup analysis further indicated increased prevalence of anti‐HEV IgG in the swine workers in Asia (PR = 1.49, 95% CI: 1.35–1.64), in Europe (PR = 1.93, 95% CI: 1.49–2.50) and in all five swine‐related occupations, including swine farmers, butchers, meat processors, pork retailers and veterinarians (PR ranged between 1.19 and 1.75). In summary, swine workers have a relatively higher prevalence of past HEV infection, and this finding is true across swine‐related occupations, which confirms zoonotic transmission between swine and swine workers.     

Hepatitis E virus in wild rabbits and European brown hares in Germany

Recently, a change of hepatitis E from being a typical travel‐associated disease to an autochthonous zoonosis in Germany was observed. An increasing number of autochthonous infections with the hepatitis E Virus (HEV) have been recognized in developed countries. Venison from wild boar is already known to be a potential source of infection, if not prepared properly by the consumer. In Germany, certain wild animals are known to be a reservoir for HEV. However, current information is missing about European brown hares (Lepus europaeus) and wild rabbits (Oryctolagus cuniculus). Thus, a total of 833 hunting‐harvested animals (European brown hares n = 669; wild rabbits n = 164) were tested for the occurrence of HEV RNA and HEV antibodies. For this, liver and blood specimens were taken after hunts in six German federal states. HEV antibodies were found by ELISA in 2.2% (624/14) of European brown hares, but no HEV RNA was detectable by nested real‐time RT‐PCR. In contrast, a seroprevalence of 37.3% (126/47) was observed for wild rabbits, and 17.1% (164/28) of the samples were HEV RNA positive. Genomic analysis revealed that these partial sequences clustered within the rabbit clade of HEV‐3 genotype. In addition, one rabbit sequence segregated into subtype 3g of HEV‐3. Highest seroprevalences for hares and rabbits were detected in the federal states of Bavaria and of Schleswig‐Holstein, respectively. Comparing urban, rural and insular areas, the highest seroprevalence was shown for wild rabbits in rural areas and for European brown hares on the northern island Fehmarn. This study provides evidence that European brown hares and wild rabbits from Germany can be infected with HEV. The different prevalences indicate that wild rabbits are a potential reservoir for HEV in Germany, whereas European brown hares seem to be only of minor importance for the epidemiology of HEV.     

Hepatitis E virus and related viruses in wild,domestic and zoo animals: A review

Hepatitis E is a human disease mainly characterized by acute liver illness, which is caused by infection with the hepatitis E virus (HEV). Large hepatitis E outbreaks have been described in developing countries; however, the disease is also increasingly recognized in industrialized countries. Mortality rates up to 25% have been described for pregnant women during outbreaks in developing countries. In addition, chronic disease courses could be observed in immunocompromised transplant patients. Whereas the HEV genotypes 1 and 2 are mainly confined to humans, genotypes 3 and 4 are also found in animals and can be zoonotically transmitted to humans. Domestic pig and wild boar represent the most important reservoirs for these genotypes. A distinct subtype of genotype 3 has been repeatedly detected in rabbits and a few human patients. Recently, HEV genotype 7 has been identified in dromedary camels and in an immunocompromised transplant patient. The reservoir animals get infected with HEV without showing any clinical symptoms. Besides these well‐known animal reservoirs, HEV‐specific antibodies and/or the genome of HEV or HEV‐related viruses have also been detected in many other animal species, including primates, other mammals and birds. In particular, genotypes 3 and 4 infections are documented in many domestic, wildlife and zoo animal species. In most cases, the presence of HEV in these animals can be explained by spillover infections, but a risk of virus transmission through contact with humans cannot be excluded. This review gives a general overview on the transmission pathways of HEV to humans. It particularly focuses on reported serological and molecular evidence of infections in wild, domestic and zoo animals with HEV or HEV‐related viruses. The role of these animals for transmission of HEV to humans and other animals is discussed.     

First detection of Hepatitis E virus (Orthohepevirus C) in wild brown rats (Rattus norvegicus) from Great Britain

In the United Kingdom, there has been an increase in the number of hepatitis E virus (HEV) infections in people annually since 2010. Most of these are thought to be indigenously acquired Orthohepevirus A genotype 3 (HEV G3), which has been linked to pork production and consumption. However, the dominant subgroup circulating in British pigs differs from that which is found in people; therefore, an alternative, potentially zoonotic, source is suspected as a possible cause of these infections. Rodents, brown rats (Rattus norvegicus) in particular, have been shown to carry HEV, both the swine HEV G3 genotype and Orthohepevirus C, genotype C1 (rat HEV). To investigate the prevalence of HEV in British rodents, liver tissue was taken from 307 rodents collected from pig farms (n = 12) and other locations (n = 10). The RNA from these samples was extracted and tested using a pan‐HEV nested RT‐PCR. Limited histopathology was also performed. In this study, 8/61 (13%, 95% CI, 5–21) of brown rat livers were positive for HEV RNA. Sequencing of amplicons demonstrated all infections to be rat HEV with 87%–92% nucleotide identity to other rat HEV sequences circulating within Europe and China (224 nt ORF‐1). Lesions and necrosis were observed histologically in 2/3 samples examined. No rat HEV RNA was detected in any other species, and no HEV G3 RNA was detected in any rodent in this study. This is the first reported detection of rat HEV in Great Britain. A human case of rat HEV infection has recently been reported in Asia, suggesting that rat HEV could pose a risk to public health.     

Hepatitis E virus in Cuba: A cross-sectional serological and virological study in pigs and people occupationally exposed to pigs

Surveillance of hepatitis E virus (HEV) in risk groups is an important strategy to monitor its circulation pattern and to timely detect changes thereof. The aims of this cross-sectional study were to estimate the prevalence of HEV infections in pigs and humans from different regions of the country, to identify risk factors for increasing anti-HEV IgG prevalence and to characterize HEV strains. The presence of anti-HEV antibodies was assessed by commercial ELISA in serum samples from the general population, farm and slaughterhouse employees, as well as pigs sampled in the three regions of Cuba from February to September 2016. Overall, individuals with occupational exposure to swine or swine products (70/248, 28.2%) were 4 times more likely to be seropositive compared to the general population (25/285, 8.7%; OR: 4.18; p < .001). Within the risk group, risk factors included age, number of years working in a professional activity with direct exposure to swine, geographic region and distance between residence and closest professional swine setting, while wearing gloves had a protective effect. Prevalence of total anti-HEV antibodies in swine was 88.2% (165/187) and HEV RNA was detected by real-time RT-PCR in 9.2% (16/173) swine stools. All HEV strains sequenced clustered within genotype 3. Some strains clearly belonged to subtype 3a, while another group of strains was related with subtypes 3b and 3 k but partial HEV sequences did not allow unequivocal subtype assignment. These findings suggest that the high HEV exposure in Cuban individuals with swine-related occupations could be due to enzootic HEV in certain regions, direct contact with infectious animals or their products as well as environmental contamination.     

Use of n-alkanes to estimate feed intake in ruminants: a meta-analysis

Precise techniques to estimate feed intake by ruminants are critical to enhance feed efficiency and to reduce greenhouse gas emissions and nutrient losses to the environment. Using a meta-analysis, we evaluated the accuracy of the n-alkane technique to predict feed intake in cattle and sheep and assessed the relationships between feed intake and fecal recovery (FR) of n-alkanes. The database was composed of 28 studies, including 129 treatments (87 and 42 for cattle and sheep, respectively) and 402 animals (232 cattle and 170 sheep) fed at troughs, from published studies. Relationships between observed (in vivo measurement) and predicted feed intake by C31:C32 and C32:C33 n-alkane pairs were evaluated by regression. Meta-regression addressed the relationships between the difference in FR of n-alkane pairs and the error in intake estimation, as well as the amount and duration of C32 n-alkane dosing. Regression of observed intake on n-alkane-based estimates revealed good relationships in cattle (adjusted R² = 0.99 for C31:C32, and adjusted R² = 0.98 for C32:C33; P < 0.0001) and in sheep (adjusted R² = 0.94 for C31:C32, and adjusted R² = 0.96 for C32:C33; P < 0.0001). FR of natural n-alkanes showed a coefficient of variation of about 15% and 16% for C31 and C33, respectively, in cattle. In sheep, the coefficient of variation was 8% and 14% for C31 and C33, respectively. The relationships between the difference of FR of n-alkane pairs and the error in feed intake estimation in cattle were characterized by an adjusted R² = 0.83 for C31:C32 (P < 0.0001) and adjusted R² = 0.93 for C32:C33 (P < 0.0001). In sheep, they were characterized by an adjusted R² = 0.69 for C31:C32 (P < 0.001) and adjusted R² = 0.76 for C32:C33 (P < 0.001). The n-alkane technique provided the reliability for estimating feed intake in cattle and sheep in barn experiments. The present meta-analysis demonstrated that without correction for differences in FR of n-alkane pairs, deviation in feed intake prediction would occur. However, further research is necessary to determine the relationship between the n-alkane dosing procedure (daily amount and duration of dosing) and FR of n-alkane.     

Cutaneous Listeriosis in a Veterinarian with the Evidence of Zoonotic Transmission – A Case Report

A case of Listeria monocytogenes skin infection in a man is presented. A 54‐year‐old male veterinary practitioner developed pustular changes on the skin of arms and hands after assisting with the delivery of a stillborn calf. Listeria monocytogenes was isolated from the skin lesions on the arms and from the bovine placenta. Listeria monocytogenes isolates were serotyped and genotyped with pulsed‐field gel electrophoresis (PFGE) to confirm the suspected transmission of the pathogen from animal to human. All isolates were of serotype 4b with identical pulsotype. To the best of our knowledge, this is the first case of cutaneous listeriosis in which the evidence for zoonotic transmission of L. monocytogenes is supported by genotyping methods.     

Hepatitis E Virus of Subtype 3a in a Pig Farm,South‐Eastern France

Hepatitis E virus (HEV) has emerged during the past decade as a causative agent of autochthonous hepatitis and is a clinical concern in Western developed countries. It has been increasingly recognized that pigs are a major reservoir of HEV of genotypes 3 and 4 worldwide and pig‐derived food items represent a potential source of infections by these viruses in humans. Hepatitis E virus RNA testing was performed here on faeces from rectal swabs sampled in 2012 from 50 3‐month‐old farm pigs from the same farm located in south‐eastern France than in a previous work conducted in 2007. Pig HEV sequences corresponding to genomic fragments of ORF2 and ORF1 genes were obtained after RT‐PCR amplification with in‐house protocols. Hepatitis E virus genotype was determined by phylogenetic analysis. Prevalence was similar to that determined 5 years earlier (68% versus 62%). Two robust phylogenetic clusters of HEV subtypes 3a and 3f were identified, and these sequences obtained in 2012 largely differ compared with those obtained in 2007. Notably, HEV sequences obtained in 2012 from a majority (62%) of the infected pigs belonged to subtype 3a, which was not previously described in France, including not being found in any of humans, pigs or wild boars. Further studies are needed to assess the circulation of HEV‐3a in pigs and humans in this country. In addition, along with previous findings, this study supports the need for increased information to the public on the risk of HEV infection through contacts with pigs or consumption of pig‐derived products in France.     

Pigs orally inoculated with swine hepatitis E virus are able to infect contact sentinels

The purpose of the present study was to explore the most likely natural route of infection of swine hepatitis E virus (HEV) by oral inoculation of pigs and to investigate the potential infection by direct contact exposure. A preliminary experiment was performed to assess the infectiousness of the bile used as source of virus. Once confirmed, 16 pigs were inoculated via oral drop with an HEV positive bile suspension containing 2 × 10⁵ genome equivalents per pig. Nine animals were kept as contact sentinels and 12 more pigs were used as negative controls. A number of pigs from the three groups were euthanized at 16, 32 and 64 days post-inoculation. From the HEV inoculated group, three pigs shed virus in faeces, two had virus RNA in bile at necropsy and two seroconverted. In the contact group, two animals showed presence of HEV RNA in bile. This study demonstrates that pigs orally inoculated with a single HEV dose got infection, although few animals had evidence of infection. Moreover, the virus was successfully transmitted to direct contact exposed pigs.     

High prevalence of Hepatitis E virus in French domestic pigs

The importance of the domestic pig reservoir for Hepatitis E virus (HEV) was assessed by estimating the seroprevalence and prevalence of HEV contaminated livers in French slaughter-aged pigs. 6565 sera and 3715 livers were randomly sampled from 186 pig farms throughout the country. Taking the sampling design into account, the farm-level seroprevalence was 65% (95% CI 57–74) and 31% (95% CI 24–38) of the slaughter-aged pigs had antibodies against HEV. The individual prevalence of HEV RNA positive livers was 4% (95% CI 2–6) and 24% (95% CI 17–31) of the farms had at least 1 positive liver. Most isolates were of genotype 3f (76.7%) with smaller amounts of 3c (18.6%) and 3e (4.6%). The high prevalence of HEV in pigs and the similarities between HEV subtypes from pigs and humans corroborates the possible zoonotic origin of some HEV autochthonous infections.     

Classification of worldwide bovine tuberculosis risk factors in cattle: a stratified approach

The worldwide status of bovine tuberculosis (bTB) as a zoonosis remains of great concern. This article reviews the main risk factors for bTB in cattle based on a three-level classification: animal, herd and region/country level. A distinction is also made, whenever possible, between situations in developed and developing countries as the difference of context might have consequences in terms of risk of bTB. Recommendations are suggested to animal health professionals and scientists directly involved in the control and prevention of bTB in cattle. The determination of Millenium Development Goals for bTB is proposed to improve the control/eradication of the disease worldwide.     

Upper respiratory infection of lactating sows with transmissible gastroenteritis virus following contact exposure to infected piglets.

Ten breeding sows were left in direct contact with their newborn piglets that had been experimentally infected with transmissible gastroenteritis (TGE) virus. All sows became infected with the virus. The sows developed fever and showed mild clinical signs of the disease for a few days. The sows excreted virus in the nasal secretion, feces, and milk during the acute febrile phase of illness. Virus was isolated from the nasal secretion of one sow as early as 20 hours after contact exposure to the infected piglets. At necropsy, the virus was more frequently isolated from the tissues of the upper respiratory tract than from small intestines; this finding indicated that the TGE coronavirus replicated in the upper respiratory tract and induced an acute respiratory infection in susceptible adult swine. Neutralizing antibody was present in the sera 8 sows after 12 to 36 days during the convalescent period. From these results, we conclude that susceptible sows in direct contact with ill piglets can become infected and by excreting virus can serve as a source of TGE virus for other susceptible pigs on the premises.     

Seroprevalence of Hepatitis E virus infection in the Irish pig population

Background

Hepatitis E is an acute viral disease of humans, occurring in explosive outbreaks in the developing world and as sporadic cases in returning travellers. Increasing recognition of indigenous transmission in Western countries suggests a zoonotic source of infection, most likely pigs. To determine if hepatitis E virus is present in Irish pigs, sera from 330 animals were examined for antibodies using a commercially available ELISA.

Findings

Antibodies were detected in 89 pigs (27%) in 13 herds (81%).

Conclusions

Hepatitis E virus is present in most Irish pig herds and in many animals within these herds.     

利用劳莱恩牛冻精改良粤北黄牛的技术研究

为提高粤北山区黄牛生长发育和产肉性能,引进世界优良肉牛品种劳莱恩牛,利用其冷冻精液发交改良粤北山区黄牛,生产的劳黄Ｆ１,其生长性能、产生能和肉用品质均比粤北山区有显著的提高。     

Cluster analysis to evaluate disease risk in periparturient dairy cattle

Predicting periparturient disease risk is of immense value to the dairy industry. Periparturient diseases are interrelated with each other; however, predicting the onset risk of these diseases has predominantly been based on a single blood parameter for a single disease. This study examined a new diagnostic method to predict the risk of periparturient diseases. We conducted cluster analysis of multiple blood constituents from 20 Holstein cattle at 1 week post‐partum, and the cattle were divided into two groups, A or B. We then compared the periparturient and early‐lactation blood constituents of these groups. Group B had significantly higher 3‐hydroxybutyric acid concentrations and were suspected to have subclinical ketosis. Group B also had significantly lower calcium concentrations, with a tendency for subclinical hypocalcemia. We also performed discriminant analysis using blood parameters at 1 week post‐partum, which grouped the population into the same two groups as the cluster analysis based on three variables: inorganic phosphorus, calcium, and either phospholipids or total cholesterol. We further showed that these discriminant functions could be used to predict the risk of periparturient disease even before parturition. Our results indicate that cluster analysis with multiple blood constituents is useful for predicting periparturient disease risks.     

猪戊型肝炎病毒大庆株DQ1全基因序列分析

本试验采用RT．-PCR的方法对戊型肝炎病毒大庆株（DQ1HEV株）的全基因进行分片段扩增，并对其两个末端采用末端快速扩增法（RACE）进行扩增、克隆，测序。与已报道的人的14株HEV的4个基因型的核苷酸和氨基酸进行比较，与IV型HEV的同源性最高。ORF1区与IV型HEV核苷酸的同源性为82．6％～83．6％．氨基酸同源性为93．5％，ORF2区与IV型HEV核苷酸的同源性为87．0％～88.4％，氨基酸同源性为95．8％～97．4％。ORF3区与IV型HEV核苷酸的同源性为94．4％～96．5％，氨基酸同源性为90.3％～96．5％。其结果表明DQ1 HEV株为IV型HEv。