Morphomolecular identification of heavy parasitic typhlitis in layer flocks: tissue response and cell-mediated reaction.

BACKGROUND: Heterakis gallinarum (H. gallinarum) is a common poultry parasite that can be found in the ceca of many gallinaceous bird species, causing minor pathology and reduced weight gain. Most infections go unnoticed in commercial flocks due to the dependence on fecal egg counts, which are prone to false-negative diagnoses. Furthermore, there is a lack of research on gastrointestinal nematodes that use molecular identification methods, which could be essential for rapid diagnosis and developing efficient control approaches. As a result, the study aimed to look at the cause of mortality in layer chickens induced by H. gallinarum in Egyptian poultry farms using morphological, ultrastructural, and molecular characterization. Histopathological, immunohistochemical, and cell-mediated immune responses from damaged cecal tissues were also examined. RESULTS: Seventy bird samples from ten-layer flocks of different breeds (Native, white, and brown layers) suffering from diarrhea, decreased egg output, and emaciation were collected. Cecal samples were collected from affected and non-affected birds and were examined for parasitic diseases using light and a scanning electron microscope. The mitochondrial cytochrome oxidase 1 (COX1) gene was used to characterize H. gallinarum. Our results showed that the collected nematodal worms were identified as H. gallinarum (male and female), further confirmed by COX1 gene amplification and sequence alignment. Gene expression analysis of the inflammatory markers in infected tissues showed a significant up-regulation of IL-2, IFN-γ, TLR-4, and IL-1ß and a significant down-regulation of the anti-inflammatory IL-10. The mRNA level of the apoptotic cas-3 revealed apoptotic activity among the H. gallinarum samples compared to the control group. CONCLUSIONS: Our results implemented the use of molecular methods for the diagnosis of Heterakis, and this is the first report showing the tissue immune response following infection in layers: upregulation of IL-1ß, IFN-γ, Il-2, and TLR-4, while down-regulation of anti-inflammatory IL-10 in cecal tissue, Cas-3 apoptotic activity and Nuclear factor-κB (NF-κB)activity with immunophenotyping of T-cells in Heterakis infected tissue.

Torsion of the caudate lobe of the liver and concurrent necrohemorrhagic typhlocolitis in a zoo-housed Patagonian mara.

Liver lobe torsion has been reported in many species, with frequent reports in rabbits. Here we describe caudate liver lobe torsion and concurrent necrohemorrhagic typhlocolitis in a Patagonian mara (syn: Patagonian cavy, Patagonian hare, Dolichotis patagonum). Following acute death, postmortem examination findings included torsion of the hepatic caudate process, which had fibrous adhesions to the pancreas indicating chronicity. The cecal apex and proximal 30 cm of colon had regionally reddened serosa and diffusely roughened and reddened mucosa with brown-red and granular luminal contents. Key histologic findings included massive necrosis of the torsed hepatic caudate lobe, consistent with infarction, necrotizing hepatitis in remaining areas of liver, necrohemorrhagic typhlocolitis, adrenocortical necrosis and hemorrhage, and renal tubular degeneration and necrosis with tubular casts. Bacterial culture of cecal contents yielded pure growth of Salmonella spp. Death was attributed to toxemia or bacteremia resulting from Salmonella spp. infection, as the hepatic lobe torsion appeared chronic. It was undetermined if the liver lobe torsion predisposed to gastrointestinal compromise and infection. Patagonian maras have some anatomical similarities to rabbits and are highly cursorial, not dissimilar to hares, Lepus spp. We speculate that these characteristics may increase the likelihood of hepatic caudate lobe torsion in this species.

Mycobacterium genavense granulomatous typhlocolitis in a horse.

A 23-y-old gelding was presented to a veterinary teaching hospital with a history of chronic, refractory diarrhea. Clinically, the horse was in poor body condition, with a thickened and corrugated large intestine identified by transcutaneous abdominal ultrasonography. At postmortem examination following euthanasia, the large colon and cecum had segmental thickening of the intestinal wall with innumerable mucosal ulcers and prominent polypoid mucosal masses. Many mesenteric and hepatic lymph nodes were enlarged. Histology revealed granulomatous and ulcerative typhlocolitis and granulomatous lymphadenitis with myriad acid-fast, variably gram-positive, intrahistiocytic bacilli that stained by immunohistochemistry for mycobacteria. Molecular testing by PCR and sequencing identified the causative agent as Mycobacterium genavense, which is an unusual presentation of infection in a horse.

Heterakis isolonche Infection Associated with Severe Nodular Typhlitis and Suspect Aberrant Pulmonary Migration in a Golden Pheasant (Chrysolopus pictus).

This case report describes the clinical, parasitologic, pathologic, and histologic characteristics of a golden pheasant (Chrysolopus pictus) with an infection of Heterakis isolonche in Mississippi. An approximately 2-yr-old golden pheasant from a flock of 8 to 10 birds was submitted to the Poultry Research and Diagnostic Laboratory in Pearl, MS, for necropsy. Clinical history indicated that three flock mates had died of unknown causes in the past. At necropsy, the submitted pheasant showed severe nodular typhlitis associated with the presence of numerous whitish small nematodes inside the cecal walls and lumen with morphologic features consistent with H. isolonche. The histologic examination showed multifocal to coalescing, nodular, granulomatous, and lymphocytic typhlitis with fibroplasia, and multiple intralesional nematodes. Furthermore, the presence of similar nematodes in the lung indicated a possible aberrant migration of Heterakis sp. to this organ. The flock was subsequently treated with an oxfendazole-containing dewormer and suffered no further losses.

Reporte de Caso- Infección por Heterakis isolonche asociada a tiflitis nodular severa y posible migración pulmonar aberrante en un faisán dorado (Chrysolopus pictus). Este informe de caso describe las características clínicas, parasitológicas, patológicas e histológicas de un faisán dorado (Chrysolopus pictus) con una infección por Heterakis isolonche en Mississippi. Un faisán dorado de aproximadamente dos años de edad de una parvada de ocho a diez aves fue remitido al Laboratorio de Investigación y Diagnóstico Avícolas en Pearl, Mississippi, para su necropsia. La historia clínica indicó que tres aves de la misma parvada habían muerto previamente por causas desconocidas. En la necropsia se observó tiflitis nodular grave asociada con la presencia de numerosos nematodos pequeños blanquecinos dentro de las paredes cecales y en el lumen con características morfológicas compatibles con H. isolonche. El examen histológico mostró tiflitis multifocal nodular coalescente, granulomatosa y linfocítica con fibroplasia y múltiples nematodos intralesionales. Además, la presencia de nematodos similares en el pulmón indicó una posible migración aberrante de Heterakis sp. a este órgano. Posteriormente, la parvada fue tratada con un antiparasitario que contenía oxfendazol y no presentó más pérdidas por mortalidad.

Typhlitis induced by Histomonas meleagridis affects relative but not the absolute Escherichia coli counts and invasion in the gut in turkeys.

Unlike in chickens, dynamics of the gut microbiome in turkeys is limitedly understood and no data were yet published in context of pathological changes following experimental infection. Thus, the impact of Histomonas meleagridis-associated inflammatory changes in the caecal microbiome, especially the Escherichia coli population and their caecal wall invasion in turkeys was investigated. Birds experimentally inoculated with attenuated and/or virulent H. meleagridis and non-inoculated negative controls were divided based on the severity of macroscopic caecal lesions. The high throughput amplicon sequencing of 16SrRNA showed that the species richness and diversity of microbial community significantly decreased in severely affected caeca. The relative abundances of operational taxonomic units belonging to Anaerotignum lactatifermentans, E. coli, and Faecalibacterium prausnitzii were higher and paralleled with a decreased abundances of those belonging to Alistipes putredinis, Streptococcus alactolyticus, Lactobacillus salivarius and Lactobacillus reuteri in birds with the highest lesion scores. Although the relative abundance of E. coli was higher, the absolute count was not affected by the severity of pathological lesions. Immunohistochemistry showed that E. coli was only present in the luminal content of caecum and did not penetrate even severely inflamed and necrotized caecal wall. Overall, it was demonstrated that the fundamental shift in caecal microbiota of turkeys infected with H. meleagridis was attributed to the pathology induced by the parasite, which only led to relative but not absolute changes in E. coli population. Furthermore, E. coli cells did not show tendency to penetrate the caecal tissue even when the intestinal mucosal barriers were severely compromised.

Eimeria ninakohlyakimovae casts NOX-independent NETosis and induces enhanced IL-12, TNF-α, IL-6, CCL2 and iNOS gene transcription in caprine PMN.

Eimeria ninakohlyakimovae represents a highly pathogenic coccidian parasite causing severe haemorrhagic typhlocolitis in goat kids worldwide. NETosis was recently described as an efficient defense mechanism of polymorphonuclear neutrophils (PMN) acting against different parasites in vitro and in vivo. In vitro interactions of caprine PMN with parasitic stages of E. ninakohlyakimovae (i. e. oocysts and sporozoites) as well as soluble oocyst antigens (SOA) were analyzed at different ratios, concentrations and time spans. Extracellular DNA staining was used to illustrate classical molecules induced during caprine NETosis [i. e. histones (H3) and neutrophil elastase (NE)] via antibody-based immunofluorescence analyses. Functional inhibitor treatments with DPI and DNase I were applied to unveil role of NADPH oxidase (NOX) and characterize DNA-backbone composition of E. ninakohlyakimovae-triggered caprine NETosis. Scanning electron microscopy (SEM)- and immunofluorescence-analyses demonstrated that caprine PMN underwent NETosis upon contact with sporozoites and oocysts of E. ninakohlyakimovae, ensnaring filaments which firmly entrapped parasites. Detailed co-localization studies of E. ninakohlyakimovae-induced caprine NETosis revealed presence of PMN-derived DNA being adorned with nuclear H3 and NE corroborating molecular characteristics of NETosis. E. ninakohlyakoimovae-induced caprine NETosis was found to be NOX-independent since DPI inhibition led to a slight decrease of NETosis. Exposure of caprine PMN to vital E. ninakohlyakimovae sporozoites as well as SOA resulted in up-regulation of IL-12, TNF-α, IL-6, CCL2 and iNOS gene transcription in stimulated PMN. Since vital E. ninakohlyakimovae-sporozoites induced caprine NETosis, this effective entrapment mechanism might reduce initial sporozoite epithelial host cell invasion during goat coccidiosis ultimately resulting in less macromeront formation and reduced merozoites I production.

Typhlitis Associated With Natural Trichuris sp. Infection in Cats.

Trichuris spp. infections can cause typhlitis or typhlocolitis in many species, but there are no published studies about its pathology in cats. Trichuris sp. infection in cats appears to be rare in most parts of the world but is frequent in some tropical and subtropical regions. The purpose of this study was to describe intestinal lesions associated with natural Trichuris sp. infections in cats of St. Kitts, West Indies. Comprehensive autopsies, histopathological assessment of small and large intestine, and total worm counts were performed in a cross-sectional study of 30 consecutive feline mortalities. Trichuris were found in 17 of 30 (57%; 95% confidence interval, 39%-74%) of the study cats with a median worm count of 11 (range, 1-170), indicating most cats had a low-intensity infection. Trichuris infection was associated with typhlitis but not consistency of feces or body condition score. In most cats examined, the typhlitis was categorized as mild (10/15, 67%) and, less frequently, moderate (2/15, 13%) or marked (3/15, 20%). The inflammatory infiltrate varied from predominantly eosinophilic (5/15, 33%) to neutrophilic (4/15, 27%), a mixture of eosinophilic and neutrophilic (2/15, 13%), a mixture of neutrophilic and lymphoplasmacytic (1/15, 7%), or a mixture of eosinophilic, neutrophilic, and lymphoplasmacytic (3/15, 20%). In some cats, surface erosions and catarrhal exudate were adjacent to adult worms. These findings are similar to those reported with low-intensity Trichuris infections in other species.

Clinical Salmonellosis in a Closed Colony of Blood Donor Cats.

An adult feline blood donor, group-housed in a closed colony with other blood donor cats in a laboratory animal facility, developed anorexia, abdominal pain, an abdominal mass effect, and hemorrhagic diarrhea. Ultimately Salmonella infection was diagnosed. The index cat and 2 additional cats in the closed colony had clinical signs consistent with Salmonella and yielded Salmonella serotype 4,12:i:- in fecal cultures. An extensive search for the source of Salmonella was unrewarding. With the implementation of individual housing and additional barrier precautions, combined with antibiotic treatment of the index case, all the cats survived and subsequently had multiple, negative Salmonella PCR test results. This case report highlights the potential for unlikely infections to occur, even in a closed colony of research animals, as well as the important role of sanitation in the elimination of this enteric pathogen.

Nodular typhlocolitis, heterakiasis, and mesenchymal neoplasia in a ring-necked pheasant ( Phasianus colchicus) with immunohistochemical characterization of visceral metastases.

A 9-y-old, male ring-necked pheasant ( Phasianus colchicus) was autopsied following euthanasia because of acute distress, recumbency, and dyspnea. The bird had experienced a protracted period of neuromuscular disease localized to the left sciatic nerve. Gross and histologic examination of the large intestine revealed intramural nodules predominantly comprised of atypical, whorling spindle cells with small cores of granulomatous inflammation centered on cross-sections of immature adult nematodes. The body structures of these metazoan organisms and clinical disease manifestation are consistent with Heterakis isolonche infection. Nodular spindle cell proliferations without granulomatous inflammation or intralesional nematodes were also found throughout the liver and lungs, suggesting metastasis from the intestine. Immunohistochemical staining of the hepatic and pulmonary tumor tissue with vimentin and S100 suggests a neurofibroblastic origin.

Ultrasonographic, endoscopic and histological appearances of the caecum in cats presenting with chronic clinical signs of caecocolic disease.

Objectives This study aimed to describe the ultrasonographic, endoscopic and histological characteristics of the caecum and ileocaecocolic junction in cats suffering from chronic clinical signs compatible with caecocolic disease. Methods Cats presenting with clinical signs suggestive of a caecocolic disease were prospectively recruited. All cats underwent an ultrasonographic examination of the caecum, ileum, colon, ileocolic lymph nodes and local mesenteric fat, in addition to comprehensive abdominal ultrasonography. This was followed by a colonoscopy with a macroscopic assessment of the caecocolic mucosa; caecocolic tissue samples were systematically collected for histologic analysis. Results Eighteen cats were included. Eleven of 18 cats had ultrasonographic abnormalities adjacent to the ileocaecocolic junction (lymphadenopathy, local steatitis) and 13/18 cats had abnormalities directly related to the junction (wall thickening, loss of wall layering). Seventeen of 18 cats had at least one ultrasonographic abnormality. Endoscopically, hyperaemia, oedema, discoloration and/or erosions were found in all cats. Each cat was classified as having mild or moderate-to-severe lesions according to endoscopic results; no classification could be established statistically for ultrasonographic results. The accentuation of the dimpled pattern tended to be inversely related to the severity of endoscopic lesion scoring. Histologically, a large proportion of cats showed typhlitis (13/16), one had lymphoma and two were normal. All cats with typhlitis also had colitis. There was only slight agreement between endoscopic and histological caecal results regarding the severity of lesions. Loss of caecal wall layering on ultrasound was found in 7/18 cats and, surprisingly, did not appear as a reliable predictor of the severity of inflammation or of malignancy; neither did local steatitis nor lymph node size. Conclusions and relevance Ultrasonography and endoscopy should not be used as the sole methods to investigate the ileocaecocolic region in cats with clinical signs suggestive of caecocolic disease. The presence of chronic clinical signs should routinely prompt histological biopsy.

Granulomatous typhlocolitis, lymphangitis, and lymphadenitis in a horse infected with Listeria monocytogenes, Salmonella Typhimurium, and cyathostomes.

A 15-year-old American Quarter horse mare was euthanized because of poor response to therapy for severe diarrhea. Significant gross findings were limited to the large intestines. The walls of the cecum and colon were thickened with widely scattered nodules in the mucosa and submucosa that extended into the enlarged colic lymph nodes. Microscopically, there was severe granulomatous typhlocolitis, lymphangitis, and lymphadenitis, with many intralesional Gram-positive, non-acid-fast coccobacilli and few cyathostomes. Intralesional bacteria were immunohistochemically and polymerase chain reaction (PCR) assay positive for Listeria monocytogenes. Concurrent infection with Salmonella enterica serovar Typhimurium was detected by PCR and culture. Infection with L. monocytogenes in horses is rare, and coinfection with Salmonella and small strongyles probably contributed to the development of granulomatous typhlocolitis.

First report of typhlitis/typhlohepatitis caused by Tetratrichomonas gallinarum in three duck species.

Two Red-breasted Mergansers (Mergus serrator), one Hooded Merganser (Lophodytes cucullatus), and one Common Eider (Somateria mollissima) from a German zoological collection died of necrotizing typhlitis/typhlohepatitis within 2 years. Using a newly established chromogenic in situ hybridization assay, numerous intralesional trophozoites of Tetratrichomonas gallinarum could be detected in formalin-fixed and paraffin-embedded tissues from the caeca and livers of the affected birds. Partial sequencing of the 18S rRNA-gene revealed two unique nucleotide sequences very similar to T. gallinarum strains isolated from avian and human hosts. One turkey kept in the same zoological collection succumbed to histomonosis (blackhead disease) confirmed with chromogenic in situ hybridization at the time of the first duck fatalities. This turkey also harboured T. gallinarum trophozoites within necrotic cell debris in the caecal lumen, which might be epidemiologically related to the T. gallinarum infections in the ducks.

Staphylococcal meningoencephalitis, nematodiasis, and typhlocolitis in a squirrel monkey (Saimiri sciureus).

BACKGROUND: Seizures were observed in a 16-year old male Guyanese squirrel monkey with a history of inappetence and weakness. METHODS AND RESULTS: Complete blood count, biochemical profile, and urinalysis indicated systemic disease. Nematode larvae were detected in the feces. Cerebrospinal fluid (CSF) analysis revealed leukocytes and gram-positive cocci. Staphylococcus aureus was isolated from the CSF. Histopathological evaluation revealed systemic lesions with inflammation and nematodes in the small and large intestine. CONCLUSION: This is the first report describing spontaneous staphylococcal CNS infection in a squirrel monkey.