Emerging interactions between diet, gastrointestinal helminth infection, and the gut microbiota in livestock.

Increasing evidence suggests that nutritional manipulation of the commensal gut microbiota (GM) may play a key role in maintaining animal health and production in an era of reduced antimicrobial usage. Gastrointestinal helminth infections impose a considerable burden on animal performance, and recent studies suggest that infection may substantially alter the composition and function of the GM. Here, we discuss the potential interactions between different bioactive dietary components (prebiotics, probiotics and phytonutrients) and helminth infection on the GM in livestock. A number of recent studies suggest that host diet can strongly influence the nature of the helminth-GM interaction. Nutritional manipulation of the GM may thus impact helminth infection, and conversely infection may also influence how the GM responds to dietary interventions. Moreover, a dynamic interaction exists between helminths, the GM, intestinal immune responses, and inflammation. Deciphering the mechanisms underlying the diet-GM-helminth axis will likely inform future helminth control strategies, as well as having implications for how health-promoting feed additives, such as probiotics, can play a role in sustainable animal production.

Morphological comparison of genetically differentiated Polymorphus cf. minutus types.

In the present study, we analyzed the morphology of three genetic types of the bird-infecting acanthocephalan Polymorphus cf. minutus (PspT1, PspT2, PspT3), mainly based on the cystacanth-stage obtained from amphipods (Gammarus fossarum, Gammarus pulex, Gammarus roeselii, Echinogammarus spp.). Males and females were pooled as there was no considerable difference between the sexes concerning the hook measurements. Additionally, we conducted a laboratory infection of one domestic duck for each Polymorphus type, to compare their performance and localization in this host species, and to obtain adult specimens for morphological comparison. The recovery rate from the ducks 4 weeks after infection was 16% for PspT1, 23.8% for PspT2, and 25% for PspT3. The adult worms were gravid, and the females contained mature eggs. Hook size did not differ considerably between cystacanths and adults of the respective type. The three Polymorphus types could be distinguished based on the cystacanth stage by a linear discriminant analysis that included hook measurements, proboscis length, proboscis width, and number of longitudinal hook rows and hooks per row. Furthermore, PspT3 was more different from PspT1 and PspT2 than the latter types from each other. Mainly the number of longitudinal hook rows differed in PspT3 from the existing descriptions of P. minutus (mainly 14 vs. mainly 16 rows). Potentially, PspT3 could be a non-indigenous parasite that was introduced with G. roeselii and that adapted to use the indigenous G. pulex as a host, while PspT2 might have been introduced to central Europe together with Echinogammarus spp.

Extraintestinal Acanthocephalan Oncicola venezuelensis (Oligacanthorhynchidae) in Small Indian Mongooses (Herpestes auropunctatus) and African Green Monkeys (Chlorocebus aethiops sabaeus).

We identified multiple extraintestinal cystacanths during routine postmortem examination of 3 small Indian mongooses and 2 African green monkeys from the Caribbean island of St. Kitts. In mongooses, cystacanths were encysted or free in the subcutaneous tissue, skeletal muscle, or peritoneal or pericardial cavities, whereas in the monkeys, they were in the cavity and parietal layer of the, tunica vaginalis, skeletal muscle, and peritoneal cavity. Morphological, histological, and molecular characterization identified these cystacanths as Oncicola venezuelensis (Acanthocephala: Oligacanthorhynchidae). There was minimal to mild lymphoplasmacytic inflammation associated with the parasite in the mongooses and moderate inflammation, mineralization, hemorrhage, and fibrosis in the connective tissue between the testis and epididymis in 1 monkey. We identified a mature male O. venezuelensis attached in the aboral jejunum of a feral cat, confirming it as the definitive host. Termites serve as intermediate hosts and lizards as paratenic hosts. This report emphasizes the role of the small Indian mongoose and African green monkey as paratenic hosts for O. venezuelensis.

The digestive tube of Piaractus brachypomus: gross morphology, histology/histochemistry of the mucosal layer and the effects of parasitism by Neoechinorhynchus sp.

The objective of the present study was to describe the histology and histochemistry of the mucosal layer of the digestive tube of Piaractus brachypomus, and the histopathology associated with parasitism by Neoechinorhynchus sp. The digestive tube of P. brachypomus consists of three macroscopically distinct portions: short, rectilinear and elastic-walled ooesophagus, J-shaped siphon stomach and a long intestine with rectilinear and curved portions, defined by patterns of villi as foregut, midgut, and hindgut. Histological and histochemical differences were observed in the mucosal layers of the different digestive tube regions, such as intense production of neutral and acidic mucous substances in the pseudostratified mucosal epithelium of the oesophagus; positive periodic acid Schiff reagent (PAS)reactions at the apex of the columnar epithelial cells of the stomach and increased intensity of histochemical reactions in the hindgut region. Neoechinorhynchus sp. was present in 85.7% of specimens examined, with a mean intensity of 7.4 ± 6.2 (±) and abundance of 6.33. Good health of the fish indicated by high relative condition factor values ( Kn ) and occurrence of only mild to moderate alteration in the mucosal layer indicated that Neoechinorhynchus sp. exhibits low pathogenicity towards P. brachypomus hosts in farming environments, with low levels of infection.

Globule Leukocytes and Other Mast Cells in the Mouse Intestine.

Only 2 major mast cell (MC) subtypes are commonly recognized in the mouse: the large connective tissue mast cells (CTMCs) and the mucosal mast cells (MMCs). Interepithelial mucosal inflammatory cells, most commonly identified as globule leukocytes (GLs), represent a third MC subtype in mice, which we term interepithelial mucosal mast cells (ieMMCs). This term clearly distinguishes ieMMCs from lamina proprial MMCs (lpMMCs) while clearly communicating their common MC lineage. Both lpMMCs and ieMMCs are rare in normal mouse intestinal mucosa, but increased numbers of ieMMCs are seen as part of type 2 immune responses to intestinal helminth infections and in food allergies. Interestingly, we found that increased ieMMCs were consistently associated with decreased mucosal inflammation and damage, suggesting that they might have a role in controlling helminth-induced immunopathology. We also found that ieMMC hyperplasia can develop in the absence of helminth infections, for example, in Treg-deficient mice, Arf null mice, some nude mice, and certain graft-vs-host responses. Since tuft cell hyperplasia plays a critical role in type 2 immune responses to intestinal helminths, we looked for (but did not find) any direct relationship between ieMMC and tuft cell numbers in the intestinal mucosa. Much remains to be learned about the differing functions of ieMMCs and lpMMCs in the intestinal mucosa, but an essential step in deciphering their roles in mucosal immune responses will be to apply immunohistochemistry methods to consistently and accurately identify them in tissue sections.

Histopathological and ultrastructural assessment of two mugilid species infected with myxozoans and helminths.

The histopathology and ultrastructure of the intestine of mullets, Liza ramada and Liza saliens, from Comacchio lagoons (northern Italy) naturally infected with myxozoans and helminths were investigated and described. Sixty-two (80.5%) of 77 mullets harboured one or more of the following parasites species: Myxobolus mugchelo (Myxozoa), Neoechinorhynchus agilis (Acanthocephala), Haplosplanchnus pachysomus and Dicrogaster contractus (Digenea). Co-occurrence of helminths with myxozoans was common. The main damage caused by digeneans was destruction of the mucosal epithelium of the villi, necrosis and degeneration of intestinal epithelial cells. More severe intestinal damage was caused by acanthocephalans which reach the submucosa layer with their proboscis. At the site of helminths infection, several mast cells (MCs), rodlet cells (RCs), mucous cells and few neutrophils and macrophages were observed in the epithelium. RCs and mucous cells exhibited discharge activity in close vicinity to the worm's tegument. M. mugchelo conspicuous plasmodia were encysted mainly in muscle and submucosa layers of the intestine. Indeed, spores of M. mugchelo were documented within the epithelial cells of host intestine and in proximity to MCs. Degranulation of the MCs near the myxozoans was very frequent.

Transcriptomics identified a critical role for Th2 cell-intrinsic miR-155 in mediating allergy and antihelminth immunity.

Allergic diseases, orchestrated by hyperactive CD4(+) Th2 cells, are some of the most common global chronic diseases. Therapeutic intervention relies upon broad-scale corticosteroids with indiscriminate impact. To identify targets in pathogenic Th2 cells, we took a comprehensive approach to identify the microRNA (miRNA) and mRNA transcriptome of highly purified cytokine-expressing Th1, Th2, Th9, Th17, and Treg cells both generated in vitro and isolated ex vivo from allergy, infection, and autoimmune disease models. We report here that distinct regulatory miRNA networks operate to regulate Th2 cells in house dust mite-allergic or helminth-infected animals and in vitro Th2 cells, which are distinguishable from other T cells. We validated several miRNA (miR) candidates (miR-15a, miR-20b, miR-146a, miR-155, and miR-200c), which targeted a suite of dynamically regulated genes in Th2 cells. Through in-depth studies using miR-155(-/-) or miR-146a(-/-) T cells, we identified that T-cell-intrinsic miR-155 was required for type-2 immunity, in part through regulation of S1pr1, whereas T-cell-intrinsic miR-146a was required to prevent overt Th1/Th17 skewing. These data identify miR-155, but not miR-146a, as a potential therapeutic target to alleviate Th2-medited inflammation and allergy.

Morphological and histochemical changes associated with massive infection by Neoechinorhynchus buttnerae (Acanthocephala: Neoechinorhynchidae) in the farmed freshwater fish Colossoma macropomum Cuvier, 1818 from the Amazon State, Brazil.

The study describes the morphological changes associated with parasitism by the intestinal acanthocephalan Neoechinorhynchus buttnerae in tambaqui juveniles Colossoma macropomum farmed in an excavated nursery, in Manaus (Amazon) in September 2013. After fish biometrics, analysis of macroscopic changes in morphology and counting of parasites, bowel fragments were fixed and submitted to histological and histochemical processing. All fish analyzed had acanthocephalans in the intestine; intestinal loops were milky white in color, with the presence of nodules with heavy parasitism. The changes in tissues that form the intestine varied according to the arrangement of the parasites: either free in the intestinal lumen or fixed by the proboscis on the organ wall. In the first case, the changes found were flaking, abrasion, compression, hypertrophy of goblet cells and disappearance of the villi on the mucosa, leukocytic cell infiltration in the submucosa, and muscle layer thickening. In the second case, in addition to these, other changes were observed as metaplasia in muscle tissue with its replacement by a loose connective tissue with severe leukocytic infiltration, edema in blood vessels, and necrotic foci. The histochemical analysis revealed that positive Alcian Blue mucosal cells (pH 2.5) were more expressive in parasitized intestines than in intestines not parasitized by N. buttnerae.

Pathology of enteric infections induced by the acanthocephalan Profilicollis chasmagnathi in Olrog's gull, Larus atlanticus, from Argentina.

Acanthocephalans can be pathogenic helminths of marine birds. Every year during the breeding season, there is variable mortality among prefledged chicks from the largest known Olrog's gull (Larus atlanticus) colony. Mortality has been associated with infection by the acanthocephalan Profilicollis chasmagnathi. Our aim was to study the role of chicks' size as a risk factor for intensity of infection and severe pathology, and to expand upon previous pathological findings reported in acanthocephalan-infected chicks. Size of the chick was associated with intensity of infection and number of intestinal perforations, which increased by 6.9% and 4.1%, respectively, for each millimetre increment in chick size. Infection was associated with inflammatory enteritis and granulomatous peritonitis. Complete intestinal perforations were observed in 85% and 97.3% of the studied chicks in 2005 and 2006, respectively, and they were observed very early during the post-hatching period. Our results show: (1) the presence of advanced pathology associated with acanthocephalan infections in chicks, beginning very early in the post-hatching period; and (2) significant increases in the intensity of infection and the associated pathology as a function of size of chicks, in dead chicks during this period.

Structure of capsule around acanthocephalan Corynosoma strumosum from uncommon paratenic hosts-lizards of two species.

Micromorphology and ultrastructure of capsule forming around acanthocephalan Corynosoma strumosum in uncommon paratenic hosts-lizards Lacerta agilis and Lacerta viridis-have been studied. Experimental infestation of the lizards by acanthocephalans obtained from naturally infested sea fishes showed that only small amount of parasites occurred in the intestine of the host was able to migrate into body cavity and to be encapsulated. Micromorphology of capsules of different ages from different species of lizards and micromorphology and ultrastructure of capsules at the age of 1.5 and 10 days appeared to be similar. In the capsule's structure cells of inflammatory rank were prevailing: mononuclear and multinuclear macrophages, eosinophils, and basophils. Fibroblasts were not numerous and were located only in the outer part of a capsule; exocellular collagen fibers were absent. Inflammatory character of capsule confirms the idea that lizards are unsuitable paratenic hosts for corynosomes.

Endoparasite diversity and liver alterations in Hoplerythrinus unitaeniatus and Cichlasoma bimaculatum in a quilombola area in Maranhão, Brazil.

Our aim was to assess endoparasite diversity and liver alterations in Hoplerythrinus unitaeniatus (jeju) and Cichlasoma bimaculatum (acará preto) in a quilombola area in Maranhão, Brazil. For this, 21 H. unitaeniatus and 21 C. bimaculatum were caught in a natural environment and transported to a laboratory. After these had been euthanized, endoparasites were collected and identified. Liver alterations were evaluated histological analysis based on the severity of each lesion: stage I, organ functioning not compromised; stage II, more severe lesions that impair normal functioning of the organs; and stage III, very severe and irreversible lesions. Among the fish evaluated, 71.43% H. unitaeniatus and 61.90% C. bimaculatum were parasitized. Contracaecum sp. was found in both species; while acanthocephalans, only in H. unitaeniatus. The alterations were vacuolization, nucleus in the cell periphery, deformation of the cell outline, melanomacrophage center, hyperemia, cytoplasmic degeneration and nuclear vacuolization. Through calculating a histological alteration index, it was found that 26.19% of the specimens presented lesions in stage I; 38.09% lesions in stage II and 9.52% lesions in stage III. It was concluded that there is high prevalence of Contracaecum sp. and that the liver lesions may be adaptive responses by the fish to endoparasitic infection.

Intestinal immune response of Silurus glanis and Barbus barbus naturally infected with Pomphorhynchus laevis (Acanthocephala).

Immunopathological and ultrastructural studies were conducted on the intestine of barbel Barbus barbus and sheatfish Silurus glanis that were naturally infected with the acanthocephalan Pomphorhynchus laevis. Enteric helminths often cause inflammation of the digestive tract, inducing the recruitment of different types of immune cells at the site of infection. The results of our study clearly demonstrated that mast cells (MC) were the dominant immune cells which occur at the site of inflammation in both hosts. MC were associated with fibroblasts and were found in close proximity to, and inside, the capillaries of the intestine, thus, migration of mast cells via the bloodstream was suggested. Significant degranulation of MC was present. Immunohistochemical staining revealed met-enkephalin and serotonin (5-HT) in intestinal MC of both uninfected and infected barbel and the absence of the antimicrobial peptides piscidin 3 and piscidin 4 in both species. Data are discussed with respect to host immune response to an intestinal helminth and compared with other host-parasite systems.

Fine structure of Longicollum pagrosomi (Acanthocephala: Pomphorhynchidae) and intestinal histopathology of the red sea bream, Pagrus major, infected with acanthocephalans.

The results described the structure of Longicollum pagrosomi and histopathological characters of the intestine of the red sea bream, Pagrus major, infected with acanthocephalans, using the light and electron microscopes. Among the six samples of P. major, L. pagrosomi was identified in the posterior intestine of five fish samples. Adult L. pagrosomi (total length, 8-27 mm) is divided into the presoma (proboscis, anterior neck, and posterior neck) and metasoma (trunk). The proboscis had vertically arranged hooks (40 µm in length), with ten hooks per row, and the septum was observed between the posterior neck and trunk. The tegument thickness of the proboscis was approximately 15 µm, and it was composed of thin, circular muscle fibers. The outer fibrous membrane was approximately 1 µm, and the connective tissue layer was approximately 35 µm in thickness in the anterior neck. The tegument of the posterior neck enclosed the cephalic ganglion and had longitudinal and vertical muscle fibers, and the tegument thickness was approximately 45 µm. The tegument of the body, which was approximately 1 mm in thickness, was composed primarily of muscle and collagen fibers, and the structure of the tegument was different, depending on the body region. The acanthocephalans had ovaries and oval-shaped eggs with an eggshell (77.5 × 17.1 µm), floating within the body cavity of the trunk. In the infected posterior intestine of P. major, the presoma and the anterior part of the metasoma of L. pagrosomi passed through the intestinal wall and infected the intestinal tissue, perforating the loose connective tissue. In the inflammatory connective tissue, collagen and muscle fibers were fragmented and revealed partial necrosis. Lipid drops and eosinophilic granular cells aggregated in the connective tissue of the tissue capsule. In the vicinity of the acanthocephalan, the mucosal epithelia contained hypertrophied nuclei, and the epithelial layer was collapsed. In an extreme case, the mucosal fold was degenerated because of pressure from the acanthocephalan.

Structure of capsule surrounding acanthocephalans Corynosoma strumosum in paratenic hosts of three species.

Morphology of capsules surrounding acanthocephalan Corynosoma strumosum in paratenic hosts (sea fishes of three species from the northern part of the Sea of Okhotsk) was studied. A thick layer of glycocalyx is formed on the surface of acanthocephalan's tegument in smelts Osmerus mordax dentex and Hypomesus olidus; the surrounding capsule is formed by fibroblasts and collagen fibers and do not include inflammatory cells. Besides fibroblasts, capsule of the sole Limanda aspera consists also of macrophages, granulocytes, "dark" cells, and once of erythrocytes that indicate obvious inflammatory response of the host's organism to invasion; glycocalyx on the surface of acanthocephalans from the sole is weakly developed. The obtained results allow considering the smelts as the most suitable paratenic hosts and the yellow-finned sole as unsuitable paratenic host for the studied acanthocephalans.

A 6-year survey of pathological conditions of slaughtered animals at Zango abattoir in Zaria, Kaduna State, Nigeria.

A 6-year retrospective study (2000-2005) of animals slaughtered at the Zango abattoir in Zaria, Kaduna State, Nigeria was carried out to determine disease conditions encountered in slaughtered animals. Records kept at the abattoir were analysed. A total of 69,307 cattle, 3,820 goats and 1,763 sheep were slaughtered for the period under study. Of the 69,307 cattle slaughtered for the period, 22,459 (32.41%) were males and 46,848 (67.59%) were females, while 1,763 sheep were slaughtered comprising of 506 (28.70%) males and 1,257 (71.30%) females, and 3,820 goats made up of 1,212 (31.73%) males and 2,608 (68.27%) were females. The major disease and/or pathological conditions were helminthosis (fascioliasis, haemonchosis and paramphistomosis) 16.20%, Streptothricosis 4.15%, Pericarditis 2.20%, liver cirrhosis 2.08%, abscesses 1.04%, pneumonia 0.14%, nephritis 0.05% and Contagious Bovine Pleuropneumonia, Tuberculosis and Nocardiosis 0.01% each. Out of the 15,075 infected organs, 13,314 (88.38%) were partially salvaged while 1,751 (11.6%) whole organs were condemned. A total of 1,239 pregnant cows, 221 pregnant ewes and 637 pregnant does were slaughtered, representing a foetal wastage of 2.65% for cattle, 17.58% for sheep and 24.43% for goats. The result of this study apart from serving as an indicator of field disease condition also demonstrates cases of serious losses in production due to slaughter of pregnant animals especially for sheep and goats.

Case report: Intestinal perforation and secondary peritonitis due to Acanthocephala infection in a black-bellied pangolin (Phataginus tetradactyla).

A case of Acanthocephala infection in a 5-year-old female rehabilitated and released black-bellied pangolin (Phataginus tetradactyla), which was part of a post-release monitoring program of a pangolin research operation in the Central African Republic, is described. This represents the first report of Acanthocephala infection in this species, which lead to intestinal perforation, secondary peritonitis and ultimately to the death of the animal concerned. It is of relevance to alert the pangolin conservation and research community to a so far unreported cause of death. A case history and necropsy findings, as well as preliminary parasite identification and genetic characterization which potentially revealed a new gigantorhynchid taxon are presented.

Intestinal Histopathology due to an Acanthocephalan in Two Corvid Species from Northern Italy.

Carnivorous birds maintain parasites in the sylvatic cycle and have a role in their diffusion. The histopathology and ultrastructure of the intestine of 29 Hooded Crows (Corvus corone cornix) and 51 Eurasian Magpies (Pica pica), from the Province of Ferrara (Northern Italy), naturally infected with Sphaerirostris picae (Acanthocephala), were investigated. In both bird species, the prevalence of infection was around 10%, and the intensity of the infection in the Hooded Crows ranged from two to 12 acanthocephalans per host, whereas in the Eurasian Magpies it ranged from one to nine worms per bird. Previous records on the histopathology of acanthocephalans in birds do not provide information on the type of cells involved in the host's reaction. We aimed to gain information on the effects of acanthocephalans on the structural integrity of the birds' intestine and to describe the type of immune cells in the hosts against the parasite. Our results showed that S. picae disrupted the intestinal wall at the site of attachment by means of its neck and proboscis, and three main types of bird intestinal reactions were noticed. The most severe response of the hosts was against the proboscis because of the action of its hooks with recruitment of macrophages, giant cells, eosinophils, and heterophils. Sphaerirostris picae perforated the birds' entire intestinal wall, reaching the peritoneal visceral serosa, but it did not provoke a diffuse peritonitis.

First record of Paradiplozoon amurensis (Monogenea: Diplozoidae) in Iraq from gills of the cyprinid fish Cyprinion macrostomum.

A survey of gill parasites was conducted in 164 Cyprinion macrostomum (Cyprinidae) collected from the Tigris River at Tikreet city, Salah Al-Deen province, in central Iraq, in 2009-2010. The monogenean Paradiplozoon amurensis Akhmerov, 1974 (Diplozoidae) was observed in the gills of 26.2% of these fishes with a mean intensity of 2.3. This is the first report of P amurensis in Iraq. A description and morphometrics of P amurensis specimens are presented.

Atypical parasitic lesions in slaughtered cattle in Southern Brazil.

In this study, we described the morphological features and unusual presentations of hydatidosis, fasciolosis, and eosinophilic myositis caused by Sarcocystis species diagnosed at the slaughterhouse lines. Twenty-seven samples of atypical parasitic lesions from distinct cattle were evaluated, of which 12 corresponded to hydatidosis, 11 to fasciolosis, and 4 to eosinophilic sarcocystosis. Hydatid cysts were observed mainly in the heart, with all cases involving the left ventricle. Fasciolosis lesions involved mainly the lungs, and were characterized by a focal nodular elevated area involving the edges of the lobes. Intralesional trematodes were observed in three cases. Sarcocystosis lesions were observed in four cases, and lesions were primarily located in the skeletal and cardiac muscles. Grossly, they presented as focal or focally extensive streaks, patches, or numerous nodules that were greenish to mildly yellowish. Histologically, all the cases had intralesional ruptured cysts of Sarcocystis that were associated with severe eosinophilic myositis and myocarditis. Parasitic lesions in atypical locations and/or with aberrant responses should be promptly identified because it may mistakenly diagnosed as other potentially zoonotic diseases, such as cysticercosis and tuberculosis.