Evaluation of exhaust air dust polymerase chain reaction as a supplement method for soiled bedding sentinel monitoring in specific pathogen free mouse facility using two different individually ventilated cage racks.

Health monitoring is essential for ensuring animal health and reliable research results. Each animal facility should establish adequate health monitoring methods, and microbiological quality control should be implemented through regular health surveillance. Recently, specific pathogen free (SPF) mice have been housed in individually ventilated cage (IVC) racks in the majority of mouse facilities globally, and health monitoring is implemented using a soiled bedding sentinel (SBS). Even though SBS monitoring is a standard method, it has a limitation in that some pathogens are not sufficiently transmitted to the sentinel housed in the IVC. The exhaust air dust polymerase chain reaction (EAD PCR) method has been reported to be a reliable complementary method to SBS monitoring based on research findings. In Korea, health monitoring programs using EAD PCR have not yet been applied to laboratory animal facilities. The microbiological status of mouse colonies housed in the two IVC racks was compared using SBS and EAD PCR monitoring in our SPF mouse facility. Except for Helicobacter spp. and Staphylococcus aureus, the detection of 16 pathogens did not differ between the two methods. In the detection of Helicobacter spp., EAD PCR was found to be more sensitive than SBS. Helicobacter spp. were not found by SBS, whereas four S. aureus positive samples were detected by either SBS or EAD PCR test. According to our findings, EAD PCR can be used as a supplement to SBS monitoring. Moreover, EAD PCR can reduce the number of animals used, making it a 3R (Replacement, Reduction, Refinement)-consistent method.

Prevalence, diversity and public health implications of Helicobacter species in pet and stray dogs.

Background: It is probable that humans can acquire H. pylori and non-H. pylori Helicobacter infections via domestic animals. The prevalence and risk factors of infections of Helicobacter species in canines of Taipei city were therefore analysed in this study. Materials and methods: A total of 95 canine faecal samples were collected from different animal shelters and hospitals in Taipei city. Total DNA was extracted for semi-nested PCR detection of Helicobacter species. The PCR products were sequenced for further comparative database and phylogenetic analyses. Results: The overall prevalence of Helicobacter species in canines of Taipei city was 75.79% (72/95). Two gastric, seven enterohepatic and two unclassified Helicobacter species were identified, all of which have been implicated in the aetiology of human diseases. The predominant species detected included H. canis (27.78%), H. pylori (26.39%), H. canicola (18.06%), and H. bilis (13.89%) in decreasing order, while H. canadensis and H. typhlonius were identified for the first time in canines. The genotypes in H. pylori and H. canicola clusters grouped together, with their respective reference strains, showed a close evolutionary distance in the phylogenetic tree, indicating a common ancestry may have existed in these clusters respectively. The residential region of canines, dog living status (pet or stray) and breed (purebred or mixed-breed) are the risk factors associated with Helicobacter infections in the canines examined. Conclusion: The high prevalence of Helicobacter infections in canines highlights a potential public health risk of zoonotic transmission among dogs, humans and other animals, and therefore, the need for proper methods in controlling the transmission routes. In addition, the 16S rRNA gene amplification method was found to be useful for bacterial identification and phylogenetic analysis.

Evaluation of the Fecal Proteome in Healthy and Diseased Cheetahs (Acinonyx jubatus) Suffering from Gastrointestinal Disorders.

Fecal proteomics allows for the identification of proteins and peptides present in stools and is useful in finding possible new biomarkers for diagnosing and/or monitoring gastrointestinal (GI) disorders. In the present study, we investigated the fecal proteome in healthy and diseased cheetahs (Acinonyx jubatus). Captive individuals of this species frequently show gastrointestinal disorders characterized by recurrent episodes of diarrhea, rare episodes of vomiting and weight loss, associated with Helicobacter spp. infection. Fecal proteomic evaluation has been performed by two-dimensional electrophoresis followed by liquid chromatography-tandem mass spectrometry. In healthy cheetahs, the results showed the presence of the following proteins: collagen alpha-1 (II) chain, transthyretin, IgG Fc-binding protein, titin, dystonin, isopentenyl-diphosphate Delta-isomerase 1, sodium/potassium-transporting ATPase subunit alpha-1 and protein disulfide-isomerase A6. The presence of albumin isoforms was found only in diseased cheetahs. The present paper reports the study of the fecal proteome in the cheetah, evidences some differences between healthy and diseased patients and confirms, once again, the potential of fecal proteomics for the study of the GI environment, with promising developments regarding the identification of new diagnostic/monitoring markers.

Determinación de Helicobacter spp. en mucosa gástrica glandular de muías a través de la prueba de la actividad de la ureasa e histopatología / Determination of Helicobacter spp. in glandular gastric mucosa of mules through the test of urease activity and histopathology

RESUMEN La información sobre la presentación y los factores predisponentes del síndrome de úlcera gástrica en mulas (SUGM) es escasa en comparación con el síndrome de úlcera gástrica en equinos (SUGE) y asnales. Debido a la naturaleza multifactorial de este síndrome, la helicobacteriosis ha sido estudiada en otras especies. El objetivo de este trabajo fue establecer la presencia de Helicobacter spp. en mucosa gástrica de mulas a través de la prueba rápida de la ureasa (PRU) y de análisis histopatológico. Menos del 27% de las muestras reaccionaron a la PRU, con tiempos prolongados de reacción, y al Agar Urea (prueba de oro), con menor porcentaje de positividad. La histopatología reveló procesos inflamatorios crónicos, sin presencia de bacterias curvoespiraladas. Las PRU no fueron conclusivas en la determinación de Helicobacter spp., comportamiento similar reportado en equinos. Se requieren exámenes diagnósticos más específicos y procedimientos complementarios orientados a explorar por regiones del estómago en la consideración del número de muestras representativas.

ABSTRACT Information on the presentation and predisposing factors of Mule Gastric Ulcer Syndrome (MGUS) is scarce, compared to Equine Gastric Ulcer Syndrome (EGUS) and donkeys. Within the multifactorial nature of this syndrome, helicobacteriosis has been studied in other species. The objective of this work was to establish the presence of Helicobacter spp. in gastric mucosa of mules, through the rapid urease test (RUT) and histopathological analysis. Less than 27% of the samples reacted to RUTs, with prolonged reaction times, and Urea Agar (gold test), with a lower percentage of positivity. Histopathology revealed chronic inflammatory processes, without the presence of curved-spiral bacteria. The RUTs were not conclusive in the determination of Helicobacter spp., a similar behavior reported in horses. More specific diagnostic tests and complementary procedures are required to explore the regions of the stomach in consideration of the number of representative samples.

Presence of Helicobacter spp. in dental tartar and gastric mucosa, and its relationship with EGUS in horses from a public slaughterhouse

Background: Despite the fact that Helicobacter spp. has been detected in equine gastric mucosa, no evidence exists about this infection in Colombian horses affected by equine ulcerative gastric syndrome (EGUS), nor in dental tartar. Objective: To detect Helicobacter spp. DNA in equine gastric mucosa and dental tartar and determine the relationship between the presence of Helicobacter spp. and gastric lesions. Methods: Samples of glandular gastric mucosa and dental tartar were collected from 30 equine slaughterhouses. Macroscopic lesions of the stomachs were classified and the total DNA in all samples was extracted using a commercial extraction kit. A final-point PCR was performed using primers for amplification of a segment of 251 bp of the gene encoding the 16s rRNA region; the amplified fragments were subjected to a second PCR to determine the presence of H. pylori, the VacA gene was typified. The resulting amplicons were sequenced. Results: It was possible to amplify 16s rRNA in several samples but there was no amplification of VacA. Fragments of the sequences were compatible with H. heilmannii. The 23.3 and 10% of gastric and tartar samples were positive for 16s rRNA of Helicobacter spp., respectively. Conclusion: Although genetic material of Helicobacter spp. was found in some animals, there was no relationship with gastric lesions. It is possible that helicobacteriosis has no bearing in EGUS etiology.

Antecedentes: A pesar de que se ha detectado Helicobacter spp. en mucosa gástrica equina, no existe evidencia de esta infección en caballos criollos colombianos afectados por síndrome ulcerativo gástrico (SUGE), ni tampoco reportes en sarro dental. Objetivo: Detectar ADN de Helicobacter spp. en sarro dental y mucosa gástrica de equinos, y determinar la relación entre la presencia de la bacteria y lesiones gástricas. Métodos: Las muestras de mucosa glandular gástrica y sarro dental fueron colectadas de 30 equinos que se encontraban en planta de beneficio. Las lesiones macroscópicas fueron clasificadas y el ADN total de las muestras fue extraído utilizando un kit comercial. Se desarrolló PCR convencional usando cebadores específicos para la amplificación de un segmento de 251 pb de un gen que codifica la región 16S del ARNr; los fragmentos amplificados fueron sometidos a una segunda PCR para determinar la presencia de H. pylori mediante la amplificación del gen VacA. Los amplificados resultantes fueron secuenciados. Resultados: Fue posible amplificar 16s ARNr en varias muestras, pero no hubo amplificación de VacA. Los fragmentos de las secuencias fueron compatibles con H. heilmannii. El 23,3 y 10% de las muestras gástricas y sarro fueron positivas para 16s ARNr de Helicobacter spp., respectivamente. Conclusión: Aunque el material genético de Helicobacter spp. se encontró en algunos animales, no hubo relación con las lesiones gástricas. Es posible que la helicobacteriosis no tenga incidencia en la etiología del EGUS.

Antecedentes: Apesar do Helicobacter spp. ter sido detectado na mucosa gástrica de equinos, não há evidências dessa infecção em cavalos crioulos colombianos afetados pela síndrome ulcerativa gástrica (SUGE), ou no sarro. Objetivo: Detectar ADN de Helicobacter spp. na mucosa gástrica e do sarro dental de equinos, e determinar a relação entre a presença de Helicobacter spp. e lesões gástricas. Métodos: Amostras de mucosa gástrica glandular e sarro dental foram coletadas de 30 equinos de abatedouro, as lesões macroscópicas dos estômagos foram classificadas. Se realizou extração de ADN total em todas as amostras através de kit comercial. Realizou-se PCR ponto final, amplificando o segmento de 251 pb do gene que codifica para a região 16s ARNr; os fragmentos amplificados foram sometidos novamente a PCR para determinar a presença de H. pylori, ao tipificar o gene VacA, e seguidamente foram sequenciados. Resultados: O houve amplificação do 16s ARNr em várias amostras, mas não amplificação de VacA. Fragmentos das sequencias foram compatíveis com H. heilmannii. O 23,3 e 10% das amostras gástricas e do sarro foram positivas para 16s ARNr de Helicobacter, respectivamente. Conclusão: Embora material genético de Helicobacter spp. encontrou-se em algumos animais não houve relação com lesões gástricas. Possivelmente a helicobacterioses não tem papel relevante na etiologia da SUGE.

An exploratory study on the presence of Helicobacter heilmannii and Helicobacter billis in the feces of companion dogs.

BACKGROUND: Companion animals like dogs play an important role in the lives of many people and are often considered to be members of families, but definitely, any contact with them poses an inherent risk of transmitting zoonotic pathogens. One of these pathogens is the genus Helicobacter which is linked to many disorders in human and animal. OBJECTIVES: The aim of this study was to investigate the presence of some zoonotic species of genus Helicobacter in companion dogs. RESULTS: Through culturing in a special medium, nine samples (9%) were detected as infected (two pure and seven mixed culture). Based on multiplex-PCR, 13 samples (13%) were infected by Helicobacter spp. although none of them were infected by H. pylori. Species-specific PCR indicated that 38.5% or 5/13 of the samples were infected with H. heilmannii, while 15.45% or 2/13 of the samples were infected by H. billis. Multivariate logistic regression analysis showed that the age factor had a significant effect on Helicobacter spp. infection (odds ratio [OR] = 2.42, p = 0.01). CONCLUSION: This study revealed the negligible faecal transmission of H. pylori. Moreover, due to the detection of H. Heilmannii and H. billis in feces and their association with human gastric diseases, dog owners should be educated about the risks and transmission modes of zoonotic bacterial infections of dogs.

Effect of a Probiotic Mixture in Captive Cheetahs (Acinonyx Jubatus) with Gastrointestinal Symptoms-A Pilot Study.

Cheetahs (Acinonyx jubatus) are classified as "vulnerable" species due to the low numbers persisting in the wild. Gastrointestinal diseases are very common in this species when they are kept in captivity, in particular gastritis. Clinical signs are predominantly characterized by vomiting, diarrhea, weight loss and anorexia. In this study, we evaluated the efficacy of a multi-strain probiotic in two groups of cheetahs: Group A (n = 4)-rescued cheetahs housed at the Cheetah Conservation Fund (Otjiwarongo, Namibia); Group B (n = 9)-captive cheetahs housed in Italian zoos. Animals showed gastrointestinal signs of different severity, and were positive for Helicobacter spp., detected by PCR in stool samples. Two sachets of probiotic formulation were administered to all cheetahs once a day for 21 consecutive days. Clinical conditions (appetite loss, vomiting, stool consistency and Body Condition Score) before (T0) and after 21 days of probiotic administration (T1) were then compared using a simplified Feline Chronic Enteropathy Activity Index (FCEAI) score. A slight but not significant improvement in the scores was observed in Group A, which had mild intestinal symptoms, while a significant decrease in vomiting and stool consistency (**p < 0.01) scores was observed in Group B, which had more pronounced symptoms. Results suggest that high concentrations of live probiotics can be of help in managing gastrointestinal signs in cheetahs.

PCR-based detection of Helicobacter pylori and non-Helicobacter pylori species among humans and animals with potential for zoonotic infections.

Helicobacter species have been reported in animals, some of which are of zoonotic importance. This study aimed to detect Helicobacter species among human and animal samples using conventional PCR assays and to identify their zoonotic potentials. Helicobacter species was identified in human and animal samples by genus-specific PCR assays and phylogenetic analysis of partial sequencing of the 16S ribosomal RNA gene. The results revealed that Helicobacter species DNA was detected in 13 of 29 (44.83%) of the human samples. H. pylori was identified in 2 (15.38%), and H. bovis was detected in 4 (30.77%), whereas 7 (53.85%) were unidentified. H. bovis and H. heilmannii were prevalent among the animal samples. Phylogenetic analysis revealed bootstrapping of sequences with H. cinaedi in camel, H. rappini in sheep and humans, and Wollinella succinogenes in humans. In conclusion, the occurrence of non-H. pylori infections among human and animal samples suggested zoonotic potentials.

Biochemical investigation of an N-acetyltransferase from Helicobacter pullorum.

N-acetylated sugars are often found, for example, on the lipopolysaccharides of Gram-negative bacteria, on the S-layers of Gram-positive bacteria, and on the capsular polysaccharides. Key enzymes involved in their biosynthesis are the sugar N-acetyltransferases. Here, we describe a structural and functional analysis of one such enzyme from Helicobacter pullorum, an emerging pathogen that may be associated with gastroenteritis and gallbladder and liver diseases. For this analysis, the gene BA919-RS02330 putatively encoding an N-acetyltransferase was cloned, and the corresponding protein was expressed and purified. A kinetic analysis demonstrated that the enzyme utilizes dTDP-3-amino-3,6-dideoxy-d-glucose as a substrate as well as dTDP-3-amino-3,6-dideoxy-d-galactose, albeit at a reduced rate. In addition to this kinetic analysis, a similar enzyme from Helicobacter bilis was cloned and expressed, and its kinetic parameters were determined. Seven X-ray crystallographic structures of various complexes of the H. pullorum wild-type enzyme (or the C80T variant) were determined to resolutions of 1.7 Å or higher. The overall molecular architecture of the H. pullorum N-acetyltransferase places it into the Class II left-handed-ß-helix superfamily (LßH). Taken together, the data presented herein suggest that 3-acetamido-3,6-dideoxy-d-glucose (or the galactose derivative) is found on either the H. pullorum O-antigen or in another of its complex glycoconjugates. A BLAST search suggests that more than 50 non-pylori Helicobacter spp. have genes encoding N-acetyltransferases. Given that there is little information concerning the complex glycans in non-pylori Helicobacter spp. and considering their zoonotic potential, our results provide new biochemical insight into these pathogens.

Canine Gastric Carcinomas: A Histopathological and Immunohistochemical Study and Similarities with the Human Counterpart.

Canine gastric carcinoma (CGC) affects both sexes in relatively equal proportions, with a mean age of nine years, and the highest frequency in Staffordshire bull terriers. The most common histological subtype in 149 CGC cases was the undifferentiated carcinoma. CGCs were associated with increased chronic inflammation parameters and a greater chronic inflammatory score when Helicobacter spp. were present. Understanding the molecular pathways of gastric carcinoma is challenging. All markers showed variable expression for each subtype. Expression of the cell cycle regulator 14-3-3σ was positive in undifferentiated, tubular and papillary carcinomas. This demonstrates that 14-3-3σ could serve as an immunohistochemical marker in routine diagnosis and that mucinous, papillary and signet-ring cell (SRC) carcinomas follow a 14-3-3σ independent pathway. p16, another cell cycle regulator, showed increased expression in mucinous and SRC carcinomas. Expression of the adhesion molecules E-cadherin and CD44 appear context-dependent, with switching within tumor emboli potentially playing an important role in tumor cell survival, during invasion and metastasis. Within neoplastic emboli, acinar structures lacked expression of all markers, suggesting an independent molecular pathway that requires further investigation. These findings demonstrate similarities and differences between dogs and humans, albeit further clinicopathological data and molecular analysis are required.

Presence of Helicobacter pylori and H. suis DNA in Free-Range Wild Boars.

Helicobacter pylori (H. pylori) is a Gram-negative bacterium that infects half of the human population worldwide, causing gastric disorders, such as chronic gastritis, gastric or duodenal ulcers, and gastric malignancies. Helicobacter suis (H. suis) is mainly associated with pigs, but can also colonize the stomach of humans, resulting in gastric pathologies. In pigs, H. suis can induce gastritis and seems to play a role in gastric ulcer disease, seriously affecting animal production and welfare. Since close interactions between domestic animals, wildlife, and humans can increase bacterial transmission risk between species, samples of gastric tissue of 14 free range wild boars (Sus scrofa) were evaluated for the presence of H. pylori and H. suis using PCR. Samples from the antral gastric mucosa from two animals were PCR-positive for H. pylori and another one for H. suis. These findings indicate that these microorganisms were able to colonize the stomach of wild boars and raise awareness for their putative intervention in Helicobacter spp. transmission cycle.

Ménétrier-like disease in a Pointer with concurrent granulomatous gastritis, helicobacteriosis and leishmaniosis: a case report.

BACKGROUND: Ménétrier-like disease is a rare hypertrophic canine gastropathy, reported in only seven dogs. Clinical signs are vomiting, anorexia and weight loss. Macroscopically, giant cerebriform gastric mucosal folds are typically seen in the corpus and fundus of the stomach. Histopathologically, fundic mucous cell hyperplasia and loss of parietal and chief cells are typical. CASE PRESENTATION: A nine-year-old spayed female Pointer had a history of intermittent vomiting, marked weight loss and hypoalbuminaemia. A gastroduodenoscopy was performed three times within three months with macroscopic changes remaining the same. The gastric mucosa of the corpus, fundus and proximal antrum was markedly irregular, with cerebriform mucosal folds. In the first gastric biopsies, histopathology revealed a moderate granulomatous gastritis, with a severe manifestation of Helicobacter-like organisms. Treatment for Helicobacter spp. decreased the vomiting slightly. The dog was diagnosed with concurrent leishmaniosis; the conventional anti-Leishmania treatment decreased the vomiting moderately, the hypoalbuminaemia resolved and the dog gained weight back to a normal body condition. Granulomatous gastritis was not present in the gastric biopsies after these treatments. The dog increased vomiting when palliative treatment (maropitant citrate, ondansetron and esomeprazole) was discontinued, and thus, full-thickness biopsies of the stomach were taken and Ménétrier-like disease was diagnosed. The affected area was too large to be surgically removed; thus, palliative treatment was reinstated. The dog remained clinically well 39 months after the first clinical presentation. CONCLUSIONS: This is the first report of Ménétrier-like disease in a dog with a simultaneous manifestation of granulomatous gastritis, helicobacteriosis and leishmaniosis. The clinical signs decreased after treatment of helicobacteriosis and leishmaniosis, but vomiting remained probably as a sign of Ménétrier-like disease. Treatment options for dogs are surgical removal of the abnormal area or palliative treatment. In humans, promising results for a cure have been shown with cetuximab treatment, a human monoclonal antibody, but no canine antibody is commercially available yet. The dog here was doing well 39 months after first presentation, which is the longest reported survival time for Ménétrier-like disease with only palliative treatment in dogs. Full-thickness biopsies are preferred in macroscopic hypertrophic lesions of the stomach for better assessment of Ménétrier-like disease.

Deep-amplicon sequencing (DAS) analysis to determine the presence of pathogenic Helicobacter species in wastewater reused for irrigation.

Wastewater has become one of the most important and least expensive water for the agriculture sector, as well as an alternative to the overexploitation of water resources. However, inappropriate treatment before its reuse can result in a negative impact on the environment, such as the presence of pathogens. This poses an increased risk for environmental safety, which can subsequently lead to an increased risk for human health. Among all the emerging wastewater pathogens, bacteria of the genus Helicobacter are some of the most disturbing ones, since they are directly related to gastric illness and hepatobiliary and gastric cancer. Therefore, the aim of this study was to determine the presence of potentially pathogenic Helicobacter spp. in treated wastewater intended for irrigation. We used a next generation sequencing approach, based on Illumina sequencing in combination with culture and other molecular techniques (qPCR, FISH and DVC-FISH), to analyze 16 wastewater samples, with and without an enrichment step. By culture, one of the direct samples was positive for H. pylori. FISH and DVC-FISH techniques allowed for detecting viable Helicobacter spp., including H. pylori, in seven out of eight samples of wastewater from the tertiary effluents, while qPCR analysis yielded only three positive results. When wastewater microbiome was analyzed, Helicobacter genus was detected in 7 samples. The different molecular techniques used in the present study provided evidence, for the first time, of the presence of species belonging to the genus Helicobacter such as H. pylori, H. hepaticus, H. pullorum and H. suis in wastewater samples, even after disinfection treatment.

Opisthorchiasis-Induced Cholangiocarcinoma: How Innate Immunity May Cause Cancer.

Innate, inflammatory responses towards persistent Opisthorchis viverrini (OV) infection are likely to contribute to the development of cholangiocarcinoma (CCA), a liver cancer that is rare in the West but prevalent in Greater Mekong Subregion countries in Southeast Asia. Infection results in the infiltration of innate immune cells into the bile ducts and subsequent activation of inflammatory immune responses that fail to clear OV but instead may damage local tissues within the bile ducts. Not all patients infected with OV develop CCA, and so tumourigenesis may be dependent on multiple factors including the magnitude of the inflammatory response that is activated in infected individuals. The purpose of this review is to summarize how innate immune responses may promote tumourigenesis following OV infection and if such responses can be used to predict CCA onset in OV-infected individuals. It also hypothesizes on the role that Helicobacterspp., which are associated with liver fluke infections, may play in activation of the innate the immune system to promote tissue damage and persistent inflammation leading to CCA.

Helicobacter spp. in domestic cats: identification and relationship with anatomical and histopathological gastric changes in animals of blood group A / Helicobacter spp. em gatos domésticos: identificação e relação com alterações gástricas anatômicas e histopatológicas em animais de sangue tipo A

The aim of this study was to evaluate the presence of gastric Helicobacter-like organisms and the endoscopic and histopathological changes in domestic cats with blood type A. Samples from the stomach antrum, body and fundus were collected from 32 mixed-breed stray domestic cats using gastroscopy. Urease testing and cytological analysis were performed in fresh samples. Tissue sections were processed and stained with hematoxylin and eosin (H&E) and the Warthin-Starry (WS) silver staining methods for histopathological examination. Helicobacter spp. were detected in 100% of samples subjected to silver staining and cytological analysis, and in 96.9% of samples subjected to urease testing. In 87.5% of the cats, mononuclear inflammatory-cell infiltrates were identified. The graduation and distribution of inflammatory infiltrates in these cats revealed mild (78.1%) to moderate (9.4%) inflammatory changes in at least one gastric region. These changes were independent of the colonization score. Hyperplasia of the lymphoid follicles was detected in three cats. Cats of blood group A are often colonized by Helicobacter spp. and the macroscopic and microscopic findings are consistent with studies in domestic cats reported to date, concluding that the most common blood group in cats is not associated with high susceptibility to symptomatic gastritis.(AU)

O objetivo deste estudo foi avaliar a presença de organismos semelhantes a Helicobacter e as alterações endoscópicas e histopatológicas em estômago de gatos domésticos de sangue tipo A. Amostras de antro, corpo e fundo gástricos foram coletadas de 32 gatos, sem raça definida, não domiciliados através de gastroscopia. Teste de urease e análise citológica foram realizados em amostras frescas. Secções teciduais foram processadas e coradas com hematoxilina e eosina e pela prata pelo método de Warthin-Starry para avaliação histológica. Helicobacter spp. foi detectado em 100% das amostras submetidas às análises citológicas e coloração pela prata e em 96,9% das amostras submetidas ao teste de urease. Em 87,5% dos gatos foi identificado infiltrado inflamatório mononuclear. A graduação e distribuição do infiltrado inflamatório nestes gatos revelaram alterações leves (78,1%) a moderada (9,4%) em pelo menos uma região gástrica. Estas alterações eram independentes do escore de colonização. Hiperplasia de folículos linfoides foram detectadas em 3 gatos. Gatos do grupo sanguíneo A são frequentemente colonizados por Helicobacter spp. e os achados macro e microscópicos são consistentes com estudos em gatos domésticos realizados até a presente data. Conclui-se que o grupo sanguíneo mais comum em gatos não está associado com uma alta susceptibilidade a gastrite sintomática causada por Helicobacter spp.(AU)

An evaluation of the usefulness of invasive and non-invasive methods used to diagnose Helicobacter spp. infections in dogs.

The aim of this study was to assess the suitability of invasive and non-invasive methods used to diagnose Helicobacter spp. in the stomachs of dogs. The study was carried out on 30 dogs of both sexes and different breeds, between one and 15 years old. A histopathologic examination, a microbiological culture, a rapid urease test, a direct bacteriological preparation and a nested PCR assay were carried out. Gastric Helicobacter spp. was identified in gastric biopsy specimens from 16 (53.3%) dogs using direct bacteriological preparation, in four (13.3%) dogs based on a culture, in 23 (76.6%) dogs using the rapid urease test and in 21 (70,0%) dogs based on a histopathological assessment of the biopsy specimens. The nested PCR of the gastric biopsy specimens revealed gastric Helicobacter spp. in all the dogs (100%). A saliva PCR assay revealed gastric Helicobacter spp. in 23 (76.6%) dogs, while stool PCR revealed the bacterium in seven (23.3%) dogs. We found that invasive methods were more accurate than non-invasive methods in detecting a Helicobacter spp. infection in dogs. In addition, the nested PCR method used to evaluate the gastric mucosal biopsy specimens was the most accurate test for detecting Helicobacter spp. It was further found that the PCR-based saliva assay was the best non-invasive method for detecting Helicobacter spp. However, taking into consideration that most of the diagnostic methods used to detect this bacterium have drawbacks, at least two diagnostic methods should be used to detect Helicobacter spp. as is done in human medicine.

Zoonotic potential of Helicobacter spp.

The genus Helicobacter contains more than 35 species. Helicobacter pylori is the most important in terms of human health. Discovery of these helicobacters gives opportunity to understand the relationship between these bacteria which colonise the animal and human gut and their effect on the host. Infection with Helicobacter spp. and the associated diseases in their hosts allow us to study the pathogenic mechanisms. The potential zoonotic pathway for the transmission of Helicobacter spp. and epidemiology of this genus, deserve more attention to these emerging pathogens.

Co-infections with liver fluke and Helicobacter species: A paradigm change in pathogenesis of opisthorchiasis and cholangiocarcinoma?

Infection with the fish-borne liver fluke Opisthorchis viverrini is classified by the International Agency for Research on Cancer as a Group 1 carcinogen: definitely carcinogenic in humans. Cofactors likely contribute to bile duct cancer (cholangiocarcinoma) caused by this infection. Here we review recent findings that address the role of liver fluke associated H. pylori in hepatobiliary disease and malignancy. We hypothesize that co-infection by O. viverrini and the bacillus Helicobacter pylori is central of liver fluke infection associated cholangiocarcinoma.

Preclinical evaluation of anti-Helicobacter spp. activity of Hippocratea celastroides Kunth and its acute and sub-acute toxicity.

BACKGROUND: Hippocratea celastroides Kunth, commonly known as "cancerina", is used in Mexican Traditional Medicine for the treatment of gastric and intestinal infections, systemic and skin inflammation, injuries and gastritis. The aim of this research was to assess the anti-Helicobacter pylori activities of hydro-ethanolic root-bark extracts from Hippocratea celastroides Kunth in naturally infected dogs, after testing their acute and subacute toxicities in mice. METHODS: To determine in vivo acute toxicity, a hydro-ethanolic extract was obtained and administered orally in female and male Balb-C mice, at doses ranging from 2000 to 5000 mg/kg. For the subacute study, a hydro-ethanolic extract was given to male and female Balb-C mice at doses ranging from 200 to 2000 mg/kg body weight. The animals were observed daily over a period of 42 days for signs of toxicity. In the pre-clinical anti-Helicobacter spp. assay, 60 dogs were included. Eighteen and 19 dogs for the experimental and control groups respectively, concluded the study. The experimental treatment consisted of H. celastroides hydro-ethanolic extract and the control treatment of amoxicillin-clarithromycin-omeprazole. RESULTS: Oral LD50 (lethal dose 50) values for hydro-ethanolic extract were indeterminable at the highest tested doses. Under the subacute administration, neither mortality nor any sign of toxicity were observed when the hydro-ethanolic extract was administered. There were no significant alterations in biochemical parameters. The prevalence of Helicobacter spp. infection in dogs was 97.1 % for the experimental group and 100 % for the control group. Effectiveness was of 33.3 and 55 % in the experimental and control group respectively. The oral administration of H. celastroides was well-tolerated and safe. CONCLUSION: The root-bark of H. celastroides produced no signs of toxicity, and manifested pharmacological activity that indicated the possibility of an alternative treatment for H. pylori infection. Effectiveness is still low so it is necessary to continue research.

Helicobacteriose em leitões: imuno-histoquímica em amostras colhidas por meio de gastroscopia / Helicobacter infection in piglets: Immunohistochemical analysis in mucosal samples collected using gastroscopy

O diagnóstico da úlcera gastroesofágica (UGE) é fundamental para que se tente a recuperação dos afetados, entretanto há grande dificuldade na sua realização de forma precisa. Modernos endoscópios são ferramentas importantes nesse sentido e podem ser usados para determinar em que fase da criação a ulceração ocorre. A bactéria Helicobacter tem sido associada à UGE e gerado perdas econômicas. Tendo em vista que poucos trabalhos têm sido realizados para identificação do problema precocemente, este estudo teve como objetivo detectar as lesões relacionando-as, ou não, com a bactéria Helicobacter spp., mediante análises macroscópica e histopatológica de amostras colhidas por meio da gastroscopia. Foram utilizados 20 animais de ambos os sexos, pesando entre 22 e 26kg e com 65 dias de idade. O aparelho utilizado foi um gastrovideoscópio da marca Karl Storz, modelo 1380NKS. Foram colhidas amostras das regiões anatômicas aglandular (quadrilátero esofágico) e glandular (cárdica, fúndica e pilórica) para o teste ultrarrápido da urease e para as avaliações histopatológicas e imuno-histoquímicas. Onze animais apresentaram lesões na região aglandular à endoscopia, e microscopicamente 15 animais apresentaram paraqueratose. Em 18 animais, foram observadas alterações em pelo menos uma das três regiões glandulares. As lesões foram maiores na região cárdica, seguida da antral e da fúndica. Em relação ao teste ultrarrápido da urease, sete animais foram negativos nas quatro regiões, e 13 positivos em pelo menos uma delas. Em relação à imuno-histoquímica (IHQ), 10 animais foram negativos em todas as regiões e 10 foram positivos em pelo menos uma delas. Os achados pré-ulcerativos não demonstraram relação com o Helicobacter spp., que apresenta caráter saprofítico e oportunista confirmado pela sua imunomarcação em áreas sem lesão...

Gastroesophageal ulcer (GEU) diagnosis is fundamental for the treatment and recovery of the affected animal stock. GEU is a condition affecting animals, resulting in depletion of animal stock and subsequent economic losses. Helicobacter spp. have been associated with GEU. Modern endoscopes are important for detecting the stage of the breeding process at which the ulceration occurs. However, few studies regarding early detection of GEU have been conducted. Therefore, we aimed to identify whether GEU lesions were related to Helicobacter spp. infection, using gastroscopy as a diagnostic technique for macroscopic and histopathological analyses. Twenty piglets (both male and female) with a mean age of 65 days were included (weight, 22-26 kg). We used a Karl Storz Gastrovideoscope (model 1380NKS). Samples from nonglandular and glandular (cardia, fundus, and pylorus) regions were collected for the ultra-rapid urease test and for histopathological and immunohistochemical (IHC) evaluations. Eleven animals showed macroscopic lesions in the nonglandular region during endoscopy, and 15 animals showed parakeratosis on histological analysis. Lesions in at least 1 glandular region were observed in 18 animals. The lesions were bigger in the cardiac region, followed by those in the antrum and the fundus. Regarding the ultra-rapid urease test, 7 animals were negative in all 4 regions and 13 were positive in at least 1 region. On IHC, 10 animals were negative in all 4 regions and 10 were positive in at least 1 region. However, pre-ulcerative findings were not correlated with Helicobacter spp. infection in the present study. The positive IHC findings for Helicobacter spp in regions without ulcerative lesions suggest its saprophytic and opportunistic nature...