A mixed population of Helicobacter pylori,Helicobacter bizzozeronii and “Helicobacter heilmannii” in the gastric mucosa of a domestic cat

Background

The presence of Helicobacter within the gastric mucosa is responsible for producing pathology in many animal species, including man. Since humans have been shown to harbour many of the same bacterial species as domestic carnivores, concern over their zoonotic potential has been growing. Helicobacter pylori, a class 1 carcinogen responsible for cases of gastritis and gastric cancer in humans, produces similar pathology in pet carnivores and is considered an example of anthroponosis. The case here presented refers to a 13 year-old mixed breed spayed female cat seen at necropsy.

Findings

Stomach samples were analysed for the presence of Helicobacter spp. by cytology, histopathology and PCR. Mild mucosal atrophy was observed in the fundus and antrum, while lymphoplasmocytic infiltrates where noted in the lamina propria of the antrum. Helicobacter-like organisms were observed in the corpus and antrum, occupying gastric glands and surface mucosa. It was possible to detect Helicobacter spp., H. pylori, H. heilmannii and H. bizzozeronii in the fundus, corpus and antrum by PCR, while in the antrum PCR samples were positive for H. pylori.

Conclusions

The spayed female under study could represent either a yet un-described population of domestic cats infected with H. pylori or a case of anthroponosis.     

Lambs immunized with an inactivated variant of Anaplasma phagocytophilum

Background

Anaplasma phagocytophilum (formerly Ehrlichia phagocytophila) is an obligate intracellular bacterium causing the disease tick-borne fever (TBF) in domestic ruminants. An effective vaccine against the infection has been demanded for livestock by sheep farmers and veterinary practitioners for years.

Findings

In the present study, we immunized lambs with an inactivated suspension of 1 × 10⁸ killed A. phagocytophilum organisms mixed with adjuvant (Montanide ISA 61VG; Seppic). Twelve 9-months-old lambs of the Norwegian White Sheep breed were used. A full two-dose series of immunization was given subcutaneously to six lambs with a 4 week interval between injections. One month after the last immunization, all lambs were challenged with the homologous viable variant of A. phagocytophilum. After challenge, all lambs showed clinical responses for several days, although the immunized lambs reacted with an anamnestic response, i.e. significant reduction in infection rate and a significantly higher antibody titer.

Conclusion

Immunization with inactivated A. phagocytophilum did not protect lambs TBF.     

Clear distinction between Burkholderia mallei and Burkholderia pseudomallei using fluorescent motB primers

Background

A frame-shift mutation in the flagellum motor gene motB coding for the chemotaxis MotB protein of Burkholderia mallei has been utilized to design a conventional duplex PCR assay with fluorescent labelled primers.

Findings

Species specificity was tested with a panel of 13 Burkholderia type strains. A total of 41 B. mallei field strains, 36 B. pseudomallei field strains, and 1 B. thailandensis field strain from different geographic regions were tested and correctly identified. Testing of 55 non-Burkholderia bacterial species revealed 100% specificity of the assay. The minimum detection limit was 1 pg DNA or 160 GE for B. mallei and 130 GE for B. pseudomallei, respectively.

Conclusions

This assay enables the clear distinction between B. mallei and B. pseudomallei/B. thailandensis.

Electronic supplementary material

The online version of this article (doi:10.1186/s13028-015-0104-4) contains supplementary material, which is available to authorized users.     

Yersinia enterocolitica in sheep - a high frequency of biotype 1A

Background

Pigs are regarded as the main reservoir for human pathogenic Yersinia enterocolitica, which is dominated by bioserotype 4/O:3. Other animals, including sheep, have occasionally been reported as carriers of pathogenic strains of Y. enterocolitica. To our knowledge, this is the first study performed in the Nordic countries in which the presence of Y. enterocolitica in sheep is investigated.

Methods

Tonsils and faecal samples collected from sheep slaughtered on the island Gotland (Sweden) from September 2010 through January 2011 were analysed for presence of Y. enterocolitica. In an attempt to maximize recovery, several cultural strategies were applied. Various non-selective media were used and different temperatures and durations of the enrichment were applied before subculturing on Cefsulodin Irgasan Novobiocin (CIN) agar. Presumptive Y. enterocolitica colonies were subjected to urease, API 20E and agglutination test. Yersinia enterocolitica isolates were biotyped, serotyped, and tested for pathogenicity using a TaqMan PCR directed towards the ail-gene that is associated with human pathogenic strains of Y. enterocolitica.

Results

The samples collected from 99 sheep yielded 567 presumptive Y. enterocolitica colonies. Eighty urease positive isolates, from 35 sheep, were identified as Y. enterocolitica by API 20E. Thirty-four of 35 further subtyped Y. enterocolitica isolates, all from faecal samples, belonged to biotype 1A serotype O:5, O:6. O:13,7 and O:10. One strain was Yersinia mollaretii serotype O:62. No human pathogenic strains of Y. enterocolitica were found in the investigated sheep. Other species identified were Y. kristensenii (n = 4), Y. frederiksenii/intermedia (n = 3), Providencia rettgeri (n = 2), Serratia marcescens (n = 1) and Raoultella ornithinolytica (n = 1).

Conclusions

This study does not support the hypothesis that sheep play an important role in transmission of the known human pathogenic Y. enterocolitica in the studied geographical region. However, because there are studies indicating that some strains of Y. enterocolitica biotype 1A may cause disease in humans, the relative importance of sheep as carriers of human pathogenic strains of Y. enterocolitica remains unclear. Tonsils do not appear to be favourable sites for Y. enterocolitica biotype 1A in sheep.     

Majority of T. gondii seropositive chickens (Gallus domesticus) in Central Ethiopia carries the infective parasite

Background

The prevalence of Toxoplasma gondii in free range chickens is a good indicator of the prevalence of T. gondii oocysts in the environment. The aim of this study was to isolate T. gondii parasites from heart and brain of seropositive free range (FR) chickens.

Findings

Isolation of T. gondii from pooled heart and brain of 41 direct agglutination test (DAT) positive (≥1:40) free range chickens (Gallus domesticus) was carried out by bioassay in mice. T. gondii specific antibodies in mice were assayed by DAT and microscopy was employed for detection and enumeration of brain tissue cysts. Overall, bioassay was positive in 29 (70.7%) chicken samples. T. gondii tissue cysts were isolated from 59% (24/41) of bioassayed chickens: from 2 of 7 chickens with a titer of 1: ≤ 60, 2 of 5 with titer 1: 180, 6 of 8 with titer 1: 540, 10 of 15 with titer 1: 1620, 1 of 2 with titer 1: 6000, 2 of 3 with titer 1:18000, 1 of 1 with titer 1:54000. None of the isolates was pathogenic for mice. Tissue cysts were detected from 61% of seropositive mice (DAT ≥ 1:40). Generally, tissue cyst counts per brain of mouse were low (mean: 132.7 ± 84.4; range: 47–352).

Conclusions

Majority of T. gondii seropositive chickens (Gallus domesticus) in Central. Ethiopia carries the infective parasite. Tissues from the free range chicken might be a source infection for animals and humans.     

Molecular typing of Salmonella enterica serovar Enteritidis isolates from food-producing animals in Japan by multilocus variable-number tandem repeat analysis: evidence of clonal dissemination and replacement

Background

Salmonella enterica serovar Enteritidis is a zoonotic pathogen. Human infections are associated with contaminated eggs and egg products. In Japan, since 1989, the incidence of food-borne disease caused by S. Enteritidis has increased and a pandemic has occurred; however, little is known about changes that occurred before and after this pandemic event in the dominant lineage of isolates from food-producing animals. This study aimed to determine the S. Enteritidis lineages in Japan over the last few decades by using multilocus variable-number tandem repeat analysis (MLVA).

Findings

MLVA was used to analyse 79 S. Enteritidis isolates collected from chickens (n = 63), cattle (n = 12), pigs (n = 2), and goats (n = 2) during 1975–2009. The S. Enteritidis isolates showed 14 different MLVA allele combinations, which were classified into two major clusters (A and C) and a minor cluster (B). All the 62 isolates in cluster A were isolated after 1988, whereas 13 of the 17 isolates belonging to cluster B and C were isolated before 1989.

Conclusions

The MLVA results showed that cluster C was predominant before 1989, and isolates in cluster A disseminated since 1989 and replaced the previous dominant clone, suggesting that isolates of cluster A originated from imported S. Enteritidis infection.     

Antimicrobial susceptibility of porcine Brachyspira hyodysenteriae and Brachyspira pilosicoli isolated in Sweden between 1990 and 2010

Background

The anaerobic spirochetes Brachyspira hyodysenteriae and Brachyspira pilosicoli cause diarrheal diseases in pigs. Their fastidious nature has hampered standardization of methods for antimicrobial susceptibility testing. For monitoring of antimicrobial susceptibility wild type cutoff values are needed to define where the wild type distribution of MICs ends and no approved cutoffs are available for Brachyspira spp. In this study antimicrobial susceptibility data for both species (in total 906 isolates) were compiled and analyzed and wild type cut off values for B. hyodysenteriae proposed.

Methods

The MICs of tiamulin, valnemulin, tylosin, tylvalosin, doxycycline and lincomycin were determined by broth dilution in brain heart infusion broth supplemented with 10% fetal calf serum.

Results

The compiled MICs from the broth dilution tests of the B. hyodysenteriae type strain, B78^T (ATCC® 27164^T), showed that the method yields reproducible results. In an international perspective the frequencies of isolates with decreased antimicrobial susceptibility were low among both B. hyodysenteriae and B. pilosicoli. However, in B. pilosicoli a constant level of 10-15% isolates with tiamulin MICs >4 μg/ml was detected between 2002 and 2010 and in B. hyodysenteriae a gradual increase in tiamulin MICs was seen between 1990 and 2003 although this increase has ceased during the last years. The wild type cutoff values proposed for B. hyodysenteriae are: tiamulin >0.25 μg/ml, valnemulin >0.125 μg/ml, tylosin >16 μg/ml, tylvalosin >1 μg/ml, lincomycin >1 μg/ml and doxycycline >0.5 μg/ml.

Conclusions

The broth dilution method used in this study has over the years generated tightly grouped MIC populations for the field isolates and reproducible results for the control strain B78^T and is therefore a suitable antimicrobial susceptibility test method for monitoring of Brachyspira spp. Here we propose wild type cutoff values for six antimicrobial agents for B. hyodysenteriae tested by broth dilution based on MIC distributions and the current knowledge on mechanisms of resistance in this species. There are few studies on antimicrobial resistance mechanisms and MIC distributions in B. pilosicoli but to some extent the cutoff values proposed for B. hyodysenteriae may be applicable also for monitoring of antimicrobial susceptibility in B. pilosicoli.     

Assay dependence of Brucella antibody prevalence in a declining Alaskan harbor seal (Phoca vitulina) population

Background

Brucella is a group of bacteria that causes brucellosis, which can affect population health and reproductive success in many marine mammals. We investigated the serological prevalence of antibodies against Brucella bacteria in a declining harbor seal population in Glacier Bay National Park, Alaska.

Results

Prevalence ranged from 16 to 74 percent for those tests detecting antibodies, indicating that harbor seals in Glacier Bay have been exposed to Brucella bacteria. However, the actual level of serological prevalence could not be determined because results were strongly assay-dependent.

Conclusions

This study reinforces the need to carefully consider assay choice when comparing different studies on the prevalence of anti–Brucella antibodies in pinnipeds and further highlights the need for species- or taxon-specific assay validation for both pathogen and host species.     

An evaluation of the ability of Dichelobacter nodosus to survive in soil

Background

Dichelobacter nodosus is the causative agent of footrot in sheep. The survival of the bacterium in soil is of importance for the epidemiology of the disease. The investigation evaluates the survival of D. nodosus in soil with and without added hoof powder stored under different temperatures.

Results

An experimental setup was used with bacteriological culture and real-time polymerase chain reaction (PCR), and the results indicate that the bacteria can survive in soil for longer time than previously expected. The survival time was found to be dependent on temperature and the addition of hoof powder to the soil, with the longest survival time estimated to be 24 days in soil samples with hoof powder stored at 5°C.

Conclusion

Our findings indicate that the survival time of D. nodosus and its ability to infect susceptible sheep on pasture under different climatic conditions should be studied further.     

Association between average daily gain,faecal dry matter content and concentration of Lawsonia intracellularis in faeces

Background

The objective of this study was to investigate the association between average daily gain and the number of Lawsonia intracellularis bacteria in faeces of growing pigs with different levels of diarrhoea.

Methods

A longitudinal field study (n = 150 pigs) was performed in a Danish herd from day 29 to 47 post weaning. Every third day all pigs were weighed, subjected to a clinical examination and faecal samples were obtained. Faecal samples were subjected to dry matter determination and absolute quantification by PCR for L. intracellularis and porcine circovirus type 2 (PCV2). Association between average daily gain, faecal dry matter content, numbers of L. intracellularis bacteria and PCV2 genome copies in faeces was investigated in a multilevel mixed-effects linear model.

Results

Increasing numbers of L. intracellularis log₁₀ bacteria/g faeces were significantly associated with decreasing average daily gain (P < 0.001). The association was decreasing with increasing faecal dry matter content (P < 0.01). The number of PCV2 log₁₀ copies/g faeces was not significantly associated with average daily gain of the pigs (P > 0.5).

Conclusion

The results suggest a potential application of a PCR quantifying L. intracellularis in growing pigs. Faecal dry matter content must be taken into consideration in interpretation of such test results.     

Quantification of Mycobacterium avium subspecies in pig tissues by real-time quantitative PCR

Background

Mycobacterioses in animals cause economical losses and certain Mycobacterium avium subspecies are regarded as potential zoonotic agents. The evaluation of the zoonotic risk caused by M. avium subspecies requires information about the quantities of Mycobacterium strains in infected animals. Because M. avium subspecies in pig tissues are difficult or even impossible to quantify by culturing, we tested the suitability of a culture-independent real-time quantitative PCR (qPCR) assay for this purpose.

Methods

Mycobacterial DNA was extracted from porcine tissues by a novel method and quantified by Mycobacterium genus specific qPCR assay targeting the 16S rRNA gene.

Results

The response of the qPCR assay to the amount of M. avium subspecies avium mixed with porcine liver was linear in the range of approximately log10⁵ to log10⁷Mycobacterium cells per 1 g of liver. The assay was validated with three other M. avium subspecies strains. When the assay was applied to porcine lymph nodes with or without visible lesions related to Mycobacterium avium subspecies infections, around 10⁴–10⁷ mycobacterial genomes per gram of lymph nodes were detected.

Conclusions

The qPCR assay was found to be suitable for the quantification of Mycobacterium avium subspecies in porcine lymph nodes and liver.     

Antimicrobial resistant bacteria in wild mammals and birds: a coincidence or cause for concern?

Background

The emergence and dissemination of antimicrobial resistance (AMR) is a growing concern to public and animal health. The contribution attributable to wildlife remains unclear. In this study two unrelated wildlife species herring gulls (Larus argentatus) and a hybrid deer (Cervus elaphus x Cervus nippon) were investigated for the presence of Escherichia coli expressing an AMR phenotype.

Findings

Bacterial isolates resistant to β-lactam compounds were identified in both animal species and the production of functional β-lactamase was confirmed using nitrocefin. The prevalence of resistant isolates was higher in herring gulls (87%) compared to deer (31%). Resistance to this class of antibiotic was found only in non-pathogenic E. coli in herring gulls and in both pathogenic and non-pathogenic E. coli strains in deer.

Conclusions

The presence of AMR in wildlife has implications for public health, food safety and potable water source protection among others.     

Isolation and identification of Salmonella spp. from red foxes (Vulpes vulpes) and badgers (Meles meles) in northern Italy

Background

Salmonella spp. have been isolated from a wide range of wild animals. Opportunistic wild carnivores such as red foxes (Vulpes vulpes) and badgers (Meles meles) may act as environmental indicators or as potential sources of salmonellosis in humans. The present study characterizes Salmonella spp. isolated from the intestinal contents of hunted or dead red foxes (n = 509) and badgers (n = 17) in northern Italy.

Findings

Thirty-one strains of Salmonella belonging to 3 Salmonella enterica subspecies were isolated. Fourteen different serovars of S. enterica subsp. enterica were identified, among which were serovars often associated with human illness.

Conclusions

Wild opportunistic predators can influence the probability of infection of both domestic animals and humans through active shedding of the pathogen to the environment. The epidemiological role of wild carnivores in the spread of salmonellosis needs to be further studied.     

Seasonal patterns of gastrointestinal nematode infection in goats on two Lithuanian farms

Background

This study investigated seasonal changes in naturally acquired gastrointestinal nematode (GIN) infections on two Lithuanian goat farms with different parasite control practices.

Findings

On both farms, nematode faecal egg counts (FEC) and larval cultures were obtained from 15 adult and 10 young goats at bi-weekly intervals from April 2012 to April 2013. Goats on farm A were dewormed with ivermectin (0.3 mg/kg body weight) in October/November 2012, whereas the animals on farm B were left untreated. Thirteen young goats were slaughtered in August/November 2012 and April 2013 and worm burdens in the gastrointestinal tract were enumerated. In goats from both farms, Teladorsagia, Trichostrongylus, Oesophagostomum, Chabertia and Haemonchus were the dominant GIN genera. Herbage contamination with infective third-stage larvae (L₃) peaked in July/August and resulted in high FEC in September/October. Parasitological examination at slaughter showed that Teladorsagia spp. and Haemonchus contortus survived the winter, both in the abomasal mucosa as adults and as early fourth-stage larvae (EL₄). Deworming on farm A significantly reduced FEC, especially of H. contortus, at the start of the grazing period compared with the untreated farm B (P < 0.05).

Conclusions

Goats were heavily infected with several GIN throughout the year. Strategic anthelmintic treatment during housing significantly reduced nematode egg output, in particular by H. contortus, at the start of the grazing season.     

Characterization of coagulase negative staphylococci from cases of subclinical mastitis in dairy cattle in Kampala,Uganda

Background

Coagulase negative staphylococci (CNS) are the most common pathogens leading to subclinical mastitis (SCM) in dairy cattle in Uganda. Coagulase negative staphylococci can vary between bacterial species in how they cause disease. The aim of the study was to characterize CNS, from cows with SCM in Uganda, at the species level.

Findings

Quarter milk samples (n = 166) were collected from 78 animals with SCM. Bacteriological analyses were carried out at Makerere University, Kampala, Uganda and at the National Veterinary Institute (SVA), Uppsala, Sweden. The most common pathogens found in milk samples from cows with SCM were CNS (31.7%). Two species of CNS were found, S. epidermidis (85%) and S. haemolyticus (15%). Of the CNS isolates, 16/20 (80%) were positive for β-lactamase production (β+).

Conclusions

In milk samples from cows with SCM caused by CNS, S. epidermidis was most prevalent, followed by S. haemolyticus.     

Brain microabscesses in a porcine model of Staphylococcus aureus sepsis

Background

Sepsis caused by Staphylococcus aureus often leads to brain microabscesses in humans. Animal models of haematogenous brain abscesses would be useful to study this condition in detail. Recently, we developed a model of S. aureus sepsis in pigs and here we report that brain microabscesses develop in pigs with such induced S. aureus sepsis.Twelve pigs were divided into three groups. Nine pigs received an intravenous inoculation of S. aureus once at time 0 h (group 1) or twice at time 0 h and 12 h (groups 2 and 3). In each group the fourth pig served as control. The pigs were euthanized at time 12 h (Group 1), 24 h (Group 2) and 48 h (Group 3) after the first inoculation. The brains were collected and examined histopathologically.

Results

All inoculated pigs developed sepsis and seven out of nine pigs developed brain microabscesses. The microabscesses contained S. aureus and were located in the prosencephalon and mesencephalon. Chorioditis and meningitis occurred from 12 h after inoculation.

Conclusions

Pigs with experimental S. aureus sepsis often develop brain microabscesses. The porcine brain pathology mirrors the findings in human sepsis patients. We therefore suggest the pig as a useful animal model of the development of brain microabscesses caused by S. aureus sepsis.     

In vitro gastrointestinal digestion study of two wheat cultivars and evaluation of xylanase supplementation

Background

The filamentous fungus Talaromyces versatilis is known to improve the metabolizable energy of wheat-based poultry diets thanks to its ability to produce a pool of CAZymes and particularly endo-β(1,4)-xylanases. In order to appreciate their in vivo mode of action, the supplementation effect of two of its xylanases, XynD and XynB from families GH10 and GH11 respectively, have been evaluated on two different wheat cultivars Caphorn and Isengrain, which were chosen amongst 6 varieties for their difference in non starch polysaccharides content and arabinoxylan composition.

Results

Polysaccharides digestion was followed during 6 h along the digestive tract using the TNO gastrointestinal model-1, to mimic monogastric metabolism. Polysaccharide degradation appeared to occur mainly at the jejunal level and was higher with Isengrain than with Caphorn. For both cultivars, XynD and XynB supplementation increased notably the amount of reducing end sugars into the jejuno-ileal dialysates, which has been confirmed by a valuable increase of the soluble glucose into the jejunal dialysates.

Conclusions

The amounts of arabinose and xylose into the dialysates and ileal deliveries increased consequently mainly for Caphorn, suggesting that XynD and XynB supplementation in wheat-based diet could alleviate the anti-nutritional effects of arabinoxylans by limiting the physical entrapment of starch and could increase the available metabolizable energy.     

Detection of porcine circovirus genotypes 2a and 2b in aborted foetuses from infected swine herds in the State of S?o Paulo,Brazil

Background

Porcine circovirus type 2 (PCV2) has been associated with several disease complexes, including reproductive failure. The aim of this study was to identify the subtypes of PCV2 that are associated with reproductive failure in pigs from the State of São Paulo, Brazil and to investigate co-infections with other infectious organisms.

Findings

Samples of 168 aborted foetuses or mummified foetuses from five farrow-to-finish swine farms known to be infected with PCV2 and located in the State of São Paulo were tested for PCV2 by polymerase chain reaction (PCR). Positive samples were additionally tested for porcine parvovirus (PPV), Leptospira spp. and Brucella spp. by PCR. PCV2 was detected in 18 of the samples (10.7%). PPV, Brucella spp. and Leptospira spp were found in 2, 10 and 0 cases, respectively. Eleven PCV2 strains were sequenced and determined to be either genotype 2a (n = 1) or 2b (n = 10).

Conclusions

The findings indicate that the frequency of PCV2 infections in aborted porcine foetuses from the State of São Paulo is rather low (10.7%) and that co-infection with other pathogens is common and may be involved in PCV2 associated reproductive failure. No repeatable, characteristic amino acid motifs for regions of the PCV2 capsid protein seemed to be associated with abortion in sows.