First Documentation of Nematode Dispharynx sp. in Scaled Quail.

Quail populations in the United States have been declining for several decades, and the role that parasites may be playing in this decline is not well understood. The goal of this study was to document novel parasites that inhabited the scaled quail, Callipepla squamata, of the Trans-Pecos ecoregion of Texas. To do this, quail were collected by hunter-harvest, night-netting, and funnel-trapping and were necropsied in the laboratory to determine the parasites they hosted. After analyzing 386 birds, we identified Dispharynx sp. in one of the samples. This specimen is the first to be officially documented in scaled quail.

Trypanosoma brucei colonizes the tsetse gut via an immature peritrophic matrix in the proventriculus.

The peritrophic matrix of blood-feeding insects is a chitinous structure that forms a protective barrier against oral pathogens and abrasive particles1. Tsetse flies transmit Trypanosoma brucei, which is the parasite that causes human sleeping sickness and is also partially responsible for animal trypanosomiasis in Sub-Saharan Africa. For this parasite to establish an infection in flies, it must first colonize the area between the peritrophic matrix and gut epithelium called the ectoperitrophic space. Although unproven, it is generally accepted that trypanosomes reach the ectoperitrophic space by penetrating the peritrophic matrix in the anterior midgut2-4. Here, we revisited this event using fluorescence- and electron-microscopy methodologies. We show that trypanosomes penetrate the ectoperitrophic space in which the newly made peritrophic matrix is synthesized by the proventriculus. Our model describes how these proventriculus-colonizing parasites can either migrate to the ectoperitrophic space or become trapped within peritrophic matrix layers to form cyst-like bodies that are passively pushed along the gut as the matrix gets remodelled. Furthermore, early proventricular colonization seems to be promoted by factors in trypanosome-infected blood that cause higher salivary gland infections and potentially increase parasite transmission.

Atrophic Proventriculitis Associated with Gravid Females of Tetrameres sp. (Nematoda: Tetrameridae) in the Magellanic Penguin (Spheniscus magellanicus) on the Coast of Brazil.

The present study offers the first description of proventriculitis associated with the presence of gravid female nematodes of the genus Tetrameres (Nematoda: Tetrameridae) in 3 juvenile Magellanic penguins (Spheniscus magellanicus) found dead on the coast of Brazil. This study broadens knowledge on parasites associated with these hosts and the real impact of this association.

Morphological and Molecular Characterization of Larval and Adult Stages of Eustrongylides excisus (Nematoda: Dioctophymatoidea) with Histopathological Observations.

The genus Eustrongylides includes nematodes known as the etiological agent of the "big red worm disease." The aim of this work was to identify Eustrongylides spp. larvae from fish and adults from great cormorant (Phalacrocorax carbo) sampled at Lake Trasimeno, Italy, by morphological and molecular analysis. Histopathological description of the lesions in birds was also provided. We described adults of Eustrongylides excisus for the first time in Italy, and we also linked larval stages 3 and 4 to adults. The use of molecular tools combined with the traditional taxonomy will help the identification of the species, including species inquirendae. Moreover, molecular analysis can also help to investigate the role of intermediate and paratenic hosts, to deepen the knowledge about geographical distribution of the different Eustrongylides spp. and to define the zoonotic potential of E. excisus, which has not yet been identified as causal agent of human cases.

Anatomopathological changes, quantification and distribution of Libyostrongylus spp. in regions of the proventriculus and ventriculus of naturally- and experimentally-infected ostriches.

Nematodes of the genus Libyostrongylus parasitize ostriches, causing high mortality rates. These nematodes are found in the proventriculus and ventriculus of ostriches, but little is known about their distribution and the possible anatomopathological changes they cause in the various regions of these organs. This paper describes the distribution and quantification of Libyostrongylus and pathological changes found in regions of the proventriculus and ventriculus of ostriches with high and low levels of both natural and experimental infection. Ostriches were necropsied and tissue samples from the distinct regions of both organs were analysed based on nematode counts and histopathology after staining with haematoxylin and eosin, Masson's trichrome or Alcian blue/PAS. The cranial and glandular regions of the proventriculus were the most parasitized. The ventriculus contained more nematodes in the caudal region. No macro- or microscopic pathological changes were observed in either of these organs of experimentally-infected birds. However, naturally-infected birds with high levels of infection presented proventriculus with macroscopic lesions and heterophilic infiltrates surrounding nematodes. In the glandular region of this organ, nematodes were located in the adenomeres of the secretory ducts, causing altered architecture and erosions and ulcerative lesions with damaged epithelium. Nematode eggs were found in the koilin layer of the middle and caudal regions of the ventriculus only of these birds. The pH of the regions assessed by Alcian blue/PAS staining changed from acidic in the proventriculus to more alkaline in the caudal region of the ventriculus. These data add knowledge to the biology of Libyostrongylus. RESEARCH HIGHLIGHTS The most parasitized areas were the cranial and glandular regions of the proventriculus. Naturally-infected birds with high levels of infection presented macro lesions in the proventriculus and damaged epithelium. Nematode eggs were found in the ventriculus. The proventriculus had an acidic pH, which turned alkaline towards the ventriculus.

First report of Libyostrongylus douglassii (Strongylida: Trichostrongylidae) in ostriches (Struthio camelus) from Mexico.

Nematodes of the Libyostrongylus genus are hematophagous parasites found under the ostrich's proventriculus membrane: they are frequent and can cause 50% of mortality in chicks and occasionally death in adults. With the aim of determining the presence of the Libyostrongylus genus in two private collections, one located in Ayapango, State of Mexico and the other in Amacuzac, Morelos, a total of 27 fecal samples were collected between August and December of 2016. Samples were analyzed using the flotation technique. The eggs were quantified using the McMaster technique. Positive samples were cultured to obtain infective larvae and to be identified by their morphometric characteristics. Only 18 samples collected in Ayapango were positive to Libyostrongylus douglassii. This is the first time that L. douglassii is reported in Mexico.

Proventriculitis Associated with Cryptosporidium baileyi in a Snowy Owl (Bubo scandiacus) and Its Epidemiological Investigation.

We describe an unusual case of proventriculitis associated with Cryptosporidium baileyi in a 7-wk-old snowy owl (Bubo scandiacus) chick kept at a zoo. Necropsy of this animal revealed diffuse mucosal thickening of the proventriculus. Subsequent histopathological examinations of the proventriculus showed marked ductal epithelial hyperplasia with intestinal metaplasia and severe inflammatory cell infiltration in the lamina propria and submucosa. These lesions were associated with numerous periodic-acid-Schiff-positive cryptosporidia-like protozoan parasites. Moreover, oocysts found within the lamina propria had a noticeably thicker wall and displayed Ziehl-Neelsen-positive test results. PCR sequencing analyses of the 18S rDNA, actin, and 70 kDa heat shock protein gene loci identified the protozoan to be C. baileyi, of which two novel sets of primers were designed for use with formalin-fixed paraffin-embedded tissue. An epidemiological survey was carried out at the zoo to investigate the source of infection, but all owl species surveyed proved negative for cryptosporidiosis. It is most likely that small animal vectors such as wild birds or rodents were responsible for this particular lethal case. This is the first report of C. baileyi associated with proventriculitis and also the first report of cryptosporidiosis in a raptor species in Asia.

Effects of different inoculation routes on the parasitic sites of Cryptosporidium baileyi infection in chickens.

Cryptosporidiosis is prevalent in domesticated, caged, and wild birds. Cryptosporidium baileyi, an ascendant species of avian Cryptosporidium, is an important pathogen. It causes respiratory disease in chickens, especially chickens younger than 50 days. In this study, SEM, histological, semi-quantitative PCR, and nested PCR techniques were used to explore the impact of different inoculation routes on sites of C. baileyi infection in chickens. Results showed that inoculation with sporozoites or oocysts via the rectum was an effective means of causing infection. This may provide an important reference for the development of the transfection system of C. baileyi in chickens. Numerous endogenous stages of C. baileyi were observed in the bursas of Fabricius (BF) and cloacas of chickens inoculated with sporozoites or oocysts via the rectum, but no parasite was seen in the tracheas of any of these chickens. In chickens infected with oocysts via the crop, the number of parasites in the BF was approximately 23-fold more than in the trachea. All blood samples collected after inoculation were negative for C. baileyi. These data show that C. baileyi was not transferred by blood circulation between the BF and respiratory tract. Different routes of inoculation were here found to distinctly affect sites of parasitism in chickens. These findings may facilitate further understanding of the biology of C. baileyi and efforts to control avian cryptosporidiosis.

Macroscopic lesions of the ventriculus of Rhea americana , Linnaeus, 1758 (Aves: Rheidae) naturally infected by Sicarius uncinipenis (Molin, 1860) (Nematoda: Habronematidae).

There are few studies concerning the parasites of rheas. However, parasitism is the major cause of the limited success in captive breeding of these birds. Deletrocephalus dimidiatus, Deletrocephalus cesarpintoi, Paradeletrocephalus minor, and Sicarius uncinipenis are the most prevalent nematode species affecting these birds, but the lesions caused by these parasites have not been previously reported. Four adult rheas were necropsied to determine the presence or absence of gross lesions within the gastrointestinal tract, associated with parasitic infection. Two rheas parasitized by S. uncinipenis had ulcers on the koilin layer or had parasites penetrating this layer, resulting in widespread necrosis and hemorrhagic areas, whereas the 2 nonparasitized birds did not present lesions. The degree of injury was proportional to the parasitic load found in the birds. Thus, high parasitic loads can result in necrosis of the ventriculus, which may contribute to the death of birds, resulting in economic losses in the rural production of these birds.

Diagnosis and treatment of proventricular nematodiasis in an umbrella cockatoo (Cacatua alba).

CASE DESCRIPTION: A 16-year-old female umbrella cockatoo (Cacatua alba) was referred to the University of Georgia College of Veterinary Medicine for evaluation of a 3-year seasonal history of lethargy and weight fluctuation. CLINICAL FINDINGS: Abnormalities detected via clinicopathologic analyses included mild leukocytosis, heterophilia, and lymphopenia consistent with a stress leukogram. Previous fecal examinations failed to diagnose enteric parasite infestation. Results of a fecal flotation test with Sheather sugar solution revealed spirurid eggs (Spiruroidea). Coelomic radiography revealed a widened cardiohepatic waist with increased soft tissue opacity at the level of the hepatic silhouette. The caudal thoracic and abdominal air sacs bilaterally appeared compressed against the coleomic wall. The proventriculus was increased in diameter, with a proventriculus-to-keel ratio of 1.0. Coelomic ultrasonography and positive-contrast upper gastrointestinal radiography revealed severe thickening and irregularity of the proventricular wall. The animal was anesthetized for an endoscopic examination of the upper gastrointestinal tract. Intralesional nematodes were identified on histologic examination of biopsy specimens from the proventriculus. TREATMENT AND OUTCOME: Effective fenbendazole treatment (15 mg/kg [6.8 mg/lb], PO, alternating between 5 days of treatment and 5 days of no treatment, which continued for 4 periods) was confirmed by repeated endoscopy and fecal examinations. The bird remained free of clinical signs 27 months after diagnosis and treatment. CLINICAL RELEVANCE: Antemortem diagnosis of proventricular nematodiasis has not been reported in psittacines. Spirurid nematode eggs are shed intermittently, which may lead to false-negative results on a single routine fecal examination. In this patient, radiography, endoscopy, and histologic evaluation facilitated antemortem diagnosis. This is the first report of successful treatment of this condition in psittacines.

Helminth community composition, structure, and pattern in six dove species (Columbiformes: Columbidae) of South Texas.

The helminth community composition and structure of 6 species of columbids residing in south Texas are reported and compared herein. Sixty individuals of the following species, rock pigeons (Columba livia [RP]), mourning doves (Zenaida macroura [MD]), Eurasian collared-doves (Streptopelia decaocto [ECD]), white-winged doves (Zenaida asiatica [WWD]), and common ground doves (Columbina passerina [CGD]), and 48 Inca doves (Columbina inca [ID]) were collected during the summer of 2006 and examined for helminths. Twelve helminth species were found (9 nematodes and 3 cestodes), representing 486 individuals. Nematodes numerically dominated the component community in all host species. Overall, helminth prevalence was similar among host sex within all dove species. However, prevalence of Skrjabinia bonini and Hymenolepis sp. in RPs was significantly different among host age groups (P = 0.01, P = 0.0002, respectively). Likewise, prevalence of Killigrewia delafondi was higher (P = 0.0001) in adult WWDs. Based on percent similarity and Jaccard's coefficient of community indices, helminth component communities were dissimilar, and the number of shared helminth species varied among host species. Data from this study suggest that the environment surrounding preferred host habitat and foraging strategies of each host species is the driving force behind helminth component communities. This study emphasizes the importance of examining co-occurring hosts at both local and regional scales to elucidate helminth community structure and patterns.

Mixed infection with Libyostrongylus dentatus and Libyostrongylus douglassii induces a heterophilic inflammatory infiltrate in the proventriculus of ostriches.

Libyostrongylus dentatus and Libyostrongylus douglassii are haematophagous nematodes found in the proventriculus and the ventriculus of ostriches. Pathological damage leading to bird death has been attributed to L. douglassii. However, histopathology of the mixed infection has not been reported. The aim of the present work was to characterize the cellular inflammatory infiltrate found in the proventriculus of ostriches with a mixed infection. Analysis of the collected nematodes confirmed a mixed infection in the proventriculus of examined birds. Histopathological examination of the proventriculus showed an inflammatory infiltrate composed of granular cells in close proximity to the nematodes. The granulocyte infiltrate was composed mainly of heterophils identified by the lack of peroxidase and presence of fusiform granules.

Dispharynx nasuta (Nematoda:Acuariidae) infection causing proventricular lesions and death in three captive rosellas (Psittaciformes:Psittacidae).

Two adult male Crimson rosellas (Platycercus elegans) and an adult female eastern rosella (Platycercus eximius) were found dead in their cages in a private aviary in March 2009, April 2010, and February 2010, respectively, without premonitory signs. Their postmortem examination showed pectoral muscle atrophy and a distended proventriculus. The proventricular mucosal surface was covered with a gelatinous, whitish material, where a large number of nematode parasites identified as Dispharynx nasuta were present. A histologic examination revealed a hyperplastic mucosa, spirurid nematodes in the mucosal layer and inflammatory mononuclear cells in the lamina propria. This is the first report of a D. nasuta infection associated with proliferative proventriculitis and subsequent death in psittacine birds.

Molecular bases of cytoskeleton plasticity during the Trypanosoma brucei parasite cycle.

African trypanosomes are flagellated protozoan parasites responsible for sleeping sickness and transmitted by tsetse flies. The accomplishment of their parasite cycle requires adaptation to highly diverse environments. These transitions take place in a strictly defined order and are accompanied by spectacular morphological modifications in cell size, shape and positioning of organelles. To understand the molecular bases of these processes, parasites isolated from different tissues of the tsetse fly were analysed by immunofluorescence with markers for specific cytoskeleton components and by a new immunofluorescence-based assay for evaluation of the cell volume. The data revealed striking differences between proliferative stages found in the midgut or in the salivary glands and the differentiating stage occurring in the proventriculus. Cell proliferation was characterized by a significant increase in cell volume, by a pronounced cell elongation marked by microtubule extension at the posterior end, and by the production of a new flagellum similar to the existing one. In contrast, the differentiating stage found in the proventriculus does not display any increase in cell volume neither in cell length, but is marked by a profound remodelling of the posterior part of the cytoskeleton and by changes in molecular composition and/or organization of the flagellum attachment zone.

Acuaroid nematodes in the common kestrel (Falco tinnunculus) in the south of Spain.

The prevalence, intensity and abundance of acuaroid nematodes were determined in the common kestrel (Falco tinnunculus) in Andalusia, Spain. Acuaroid nematodes were present in 26/41 (63.4%) of birds examined. The most common species belonged to the genus Synhimantus subgenus Synhimantus (56%): S. (S.) laticeps (36.5%), S. (S.) robertdollfusi (24.3%) and a single specimen of a third, unknown, Synhimantus (S.) spp., unlike any other described previously (2.4%). Other species identified were Synhimantus (Dispharynx) spp. (2.4%), S. (D.) nasuta (4.8%), Desportesius spinulatus (9.7%) and Skrjabinoclava spp. (2.4%). This is the first record of these three species in F. tinnunculus, but the latter two are considered to be accidental parasites in birds of prey.

Proventricular nematodiasis in wrinkled hornbills (Aceros corrugatus).

Three immature Sunda wrinkled hornbills (Aceros corrugatus) were diagnosed postmortem with proventricular spirurid nematodiasis. Concurrent severe disseminated larval granulomatosis in other visceral organs was considered contributory to mortality in each case. Clinical signs of nematodiasis were vague but generally consisted of weight loss, anorexia, and lethargy. Frequent antemortem fecal examinations were negative for spirurid eggs. In these present cases, based on routine histopathology, both prophylactic and empirically based therapeutic anthelmintic treatments had no evident benefit in the elimination of the proventricular nematodes. Spirurid nematodiasis may be an important cause of mortality in young hornbills.

Tetrameriosis in feral pigeons from Murcia, Southeastern Spain.

A total of 165 adult feral pigeons (Columba livia) were captured in 7 different parks of the municipality of Murcia, Southeastern Spain. Birds were evaluated clinically and subsequently necropsied. All birds appeared to be in good physical condition and no apparent signs of disease were observed. However, 17.6% of the pigeons were parasitized by Tetrameres (Petrowimeres) fissispina. The nematode burden in the proventriculus ranges from 5 to 64 specimens (median: 19); the median of female and male nematodes was 17 (range: 4-54) and 3 (range: 0-12), respectively. Despite the absence of detectable signs of disease, lesions induced by T. (P.) fissispina were evident. The histopathologic analysis revealed a pressure atrophy of the proventricular glands and a light inflammatory infiltrate surrounding the papillae. Pigeons captured in parks with soil had significantly higher T. (P.) fissispina prevalence compared with those captured in paved areas.

Development of Eustrongylides ignotus (Nematoda: Dioctophmida) in domestic ducks (Anas platyrhynchos domestica (L.)).

Investigating the development of Eustrongylides ignotus in its definitive host would enable us to trace the complete life cycle of this nematode. Fourth-stage larvae isolated from naturally infected swamp eels (Monopterus albus) were used to infect domestic ducks (Anas platyrhynchos domestica [L.]). We observed that male and female worms exhibited different developmental patterns in host ducks. In males, the fourth molt occurred at day 1-2 post-infection (PI), after which they attained maturity on day 4 PI and died between day 7 and 9 PI. However, females underwent the fourth molt at day 2-4 PI, produced eggs from day 9 to 17 PI, and then degenerated and died. When compared to fourth-stage female larvae, adult females demonstrated a considerable increase in total body size with a 151% increase in average body width and a 17% increase in average body length. However, the increase in size of the male larvae was not as significant as that in females. The average body width in adult males exhibited only a 45% increase over that in the larval stage.

Differential localization of Libyostrongylus douglassii (Cobbold, 1882) Lane, 1923 and L. dentatus Hoberg, Lloyd, and Omar, 1995 (Nematoda: Trichostrongylidae) in ostrich (Struthio camelus Linnaeus, 1758) proventriculi.

Libyostrongylus is composed of 3 species, L. douglassii, L. dentatus, and L. magnus, which are differentiated based on the morphology of the adults worms. Of these, only the first 2 species have been described in Brazilian ostriches (Struthio camelus). There are very few citations regarding the occurrence of L. dentatus, having been reported only in North America and Brazil, while L. douglassii is distributed worldwide. Reports on differences in the localization of Libyostrongylus species inside the proventriculus are not available. Thirty proventriculi were collected and examined for the presence and location of the nematodes with the aid of a stereomicroscope. The collected parasites were conditioned separately, according to their sites of infection, in plates containing 0.09% saline solution. The worms were identified based on morphologic characters. All the examined proventriculi were parasitized by L. douglassii and L. dentatus. However, L. douglassii was observed under the koilin layer, while L. dentatus was inserted and attached in the koilin layer. Thus, the present study identifies different sites of infection by the 2 species of Libyostrongylus found in Brazil.

Mixed infection by Libyostrongylus douglassii and L. dentatus (Nematoda: Trichostrongylidae) in Struthio camelus (Ratites: Struthioniformes) from Brazil with further morphological characterization of adults.

The genus Libyostrongylus includes three species, L. douglassii, L. dentatus and L. magnus that occur as parasites in the proventriculus of Struthio camelus. We confirmed a mixed infection by L. douglassii and L. dentatus in farmed ostriches from the southeast of Brazil for the first time, and provided new information on some morphological characters that differentiate these species. Adult nematodes collected from the proventriculus of ostriches were observed by light and scanning electron microscopy. Morphologic characterization and morphometric analysis of the nematodes enabled the distinction of both species and corroborated results of prior studies. Specimens of L. dentatus have a buccal capsule with a prominent esophageal tooth. Furthermore, males and females of L. dentatus were larger (4954 and 9347 microm) than those of L. douglassii (3411 and 4229 microm), but measurements for most characters in both species were smaller then those previously reported. Besides, the cephalic structure based on scanning electron microscopy differs, and L. dentatus has thick lips with round papillae, whereas L. douglassii has fine lips with lengthened papillae. The confirmation of both species in South America strongly suggests that the mixed infection may be common in farmed ostriches.