Squamous cell carcinoma in a tuatara (Sphenodon punctatus)

CASE HISTORY: A mature male tuatara was presented with a history of recurrent cloacal prolapse. CLINICAL AND PATHOLOGICAL FINDINGS: The prolapsed tissue included a 12 mm diameter mass, which histologically showed cords and nodules of neoplastic epithelial cells. DIAGNOSIS: The mass was diagnosed as a squamous cell carcinoma with ulceration of the overlying stratified squamous epithelium and diffuse inflammation of the surrounding dermis. CLINICAL RELEVANCE: This case presented a rare opportunity to investigate squamous cell carcinoma in a long-lived lower vertebrate species. Future recurrence or metastasis of the mass may add useful information to the current base of knowledge of the behaviour of malignant neoplasms in reptiles.     

T cell function in tuatara (Sphenodon punctatus)

Tuatara are the sole survivors of an entire order of reptiles that thrived during the age of the dinosaurs. Therefore, knowledge of their physiology is critical to understanding the phylogeny of reptiles. Previous studies of the immune system of the tuatara did not assess T cell function. We analyzed T cell function among six captive tuatara by assessing concanavalin A (Con A), phytohemagglutinin (PHA) and mixed lymphocyte reaction (MLR) induced T cell proliferation. Peripheral blood mononuclear cells from six out of six and four out of four tuatara tested exhibited significant proliferative responses to Con A and PHA, respectively, as measured by an MTT reduction assay. A lower level of proliferation was detected in an MLR. However, Con A activated lymphocytes were not cytotoxic for a xenogeneic murine mastocytoma cell line (P815).     

Failure to detect Salmonella species in a population of wild tuatara (Sphenodon punctatus)

AIM: To assess the prevalence of faecal excretion of Salmonella serovars by wild tuatara (Sphenodon punctatus) on Stephens Island, New Zealand. METHODS: One hundred cloacal swabs obtained as part of health-screening for the translocation of adult tuatara from Stephens Island were subjected to general aerobic culture and enrichment, and cultured specifically for Salmonella spp. RESULTS: No Salmonella spp were cultured from any of the cloacal samples, which suggests that, at the 95% confidence interval, the maximum prevalence of tuatara in the island population that were shedding Salmonella spp not detected by our sample size was 1.5%. Mixed bacteria were grown from the 70 cloacal swabs cultured aerobically. A predominant organism was evident in 30 cultures, and these were identified as Hafnia alvei type 1 (n=16) and type 2 (n=7), Corynebacterium spp (n=4), Klebsiella oxytoca (n=2), and Moraxella spp (n=1). CONCLUSIONS: The absence of intestinal carriage of Salmonella spp by the tuatara sampled in this study may indicate either lack of exposure, or an innate resistance to intestinal colonisation in tuatara. The significance of the other bacteria cultured as potential pathogens to the tuatara and as zoonotic risks is also uncertain. Wildlife managers should screen translocated reptiles for Salmonella spp, and thereby avoid exposing wild and managed populations to infection.     

Mycobacterium infection in a captive-reared capercaillie (Tetrao urogallus)

One captive male capercaillie (Tetrao urogallus) was found dead on December 1993 at the breeding center of capercaillie in Catalonia, Spain. The bird was emaciated and, at necropsy, had numerous nodules of various sizes subcutaneously in the cervical region, pleura, lungs, liver, spleen, and mesentery. Microscopic examination revealed granulomatous lesions with central caseous necrosis, epithelioid cells, giant cells, and few lymphocytes in all affected organs. Numerous acid-fast bacilli were demonstrated in the tubercles with Ziehl-Nielsen stain.     

Health screening of free-ranging European brown hares (Lepus europaeus) on the German North-Sea island Pellworm

Background

A sudden decline of the European brown hare (Lepus europaeus) population in one of the best hunting districts for small game species in northern Germany, the German North-Sea island Pellworm, in the years 2007/08 following marked habitat changes led to the implementation of a thorough health assessment program of the population. 110 animals were collected during the normal hunting season in the years 2010 and 2011. A post-mortem examination and histopathological investigation was performed on all animals. Additionally, routine bacteriology of the small intestine and parasitology were carried out. Sera of hares were tested for European Brown Hare Syndrome (EBHS) by enzyme linked immunosorbent assay, and for Treponema sp. by indirect immunofluorescent test. Additional testing was performed when deemed necessary.

Results

The most striking result was a shift in the intestinal bacterial flora towards Gram-negative Enterobacteriaceae with a predominance of either Escherichia coli, or Aeromonas sp., or a high-grade double-infection with these two pathogens with subsequent catarrhal enteritis. Additionally, a marked coccidiosis, and varying infestations with the nematode Trichostrongylus retortaeformis were found. The sero-prevalence for EBHS was 78.1%, and for Treponema 43.9%.

Conclusions

The shift and decrease in diversity of the intestinal flora was the main and most consistent result found. In the authors’ opinion the change of the habitat combined with other stressors increased the animals’ sensitivity to ubiquitous bacterial species and parasites which usually would not have such fatal effects.     

Efficacy of anthelmintic treatment in captive-reared black stilts (Himantopus novaezelandiae) released to the wild

AIMS: To assess the efficacy of praziquantel (PZQ) administered to control helminths in captive-reared critically endangered black stilts (Himantopus novaezelandiae) before their release into the wild and determine the effect on their survival shortly after release.

METHODS: Of 77 captive black stilts, 37 were treated with PZQ prior to release into the wild in South Canterbury, New Zealand, in August and September 2007. Faecal helminth egg counts (FEC) were measured before and after anthelmintic treatment, and before and after release to the wild using modified faecal flotation and sedimentation methods. In addition, total helminth counts were determined in 11 of the birds that died following release, as well as four captive and 11 other wild stilts.

RESULTS: The efficacy of PZQ against trematodes was 92% and against Capillaria spp. was 34%. No trematode or Capillaria spp. eggs were detected in treated birds 1 day after treatment, but FEC increased 3–5 days after treatment. There were no differences in the total helminth counts for trematodes, cestodes or Capillaria spp. in control or treated birds (p>0.2). Survival did not differ between treatment groups in the August or September releases (p>0.4). Of control and treated birds, 11/17 (65%) and 8/14 (57%) survived to 31 days, respectively, in the August release, and 16/20 (80%) and 10/15 (67%) survived to 84 days, respectively, in the September release.

CONCLUSION: Overall, the results suggest that PZQ treatment may be an unnecessary cost and the risks of producing anthelmintic resistance, injuring the birds during processing or producing an unnecessary stress response at the time of release could exceed any likely benefits.

CLINICAL RELEVANCE: It is recommended that pre-release anthelmintic treatment of black stilts should be used only if indicated by health screening. Any treatment should incorporate annual efficacy testing to monitor the emergence of anthelmintic resistance.     

Characterization of a mannose-binding lectin from channel catfish (Ictalurus punctatus)

Mannose-binding lectin (MBL) is an important component of innate immunity capable of activating the lectin pathway of the complement system. A MBL gene was isolated from channel catfish (Ictalurus punctatus). The deduced protein contains a canonical collagen-like domain, a carbohydrate recognition domain (CRD), and a neck region similar to mammalian mannose-binding lectin. The catfish mannose-binding lectin CRD contains the EPN motif shown previously to mediate mannose specificity. The catfish mannose-binding lectin showed 30-43% identity with MBL protein sequences of rainbow trout, zebrafish, common carp, and goldfish, and 33-35% identity with sequences of mammalian species. In this study, while liver was the predominant source of mannose-binding lectin gene expression in healthy tissues, mannose-binding lectin expression in spleen rose sharply following challenge with a Gram-negative bacterium.     

First report of a Trichinella papuae infection in a wild pig (Sus scrofa) from an Australian island in the Torres Strait region

Multiple Trichinella species are reported from the Australasian region although mainland Australia has never confirmed an indigenous case of Trichinella infection in humans or animals. Wildlife surveys in high-risk regions are essential to truly determine the presence or absence of Trichinella, but in mainland Australia are largely lacking. In this study, a survey was conducted in wild pigs from mainland Australia's Cape York Peninsula and Torres Strait region for the presence of Trichinella, given the proximity of a Trichinella papuae reservoir in nearby PNG. We report the detection of a Trichinella infection in a pig from an Australian island in the Torres Strait, a narrow waterway that separates the islands of New Guinea and continental Australia. The larvae were characterised as T. papuae (Kikori strain) by PCR and sequence analysis. No Trichinella parasites were found in any pigs from the Cape York Peninsula. These results highlight the link the Torres Strait may play in providing a passage for introduction of Trichinella parasites from the Australasian region to the Australian mainland.     

Molecular and functional characterization of channel catfish (Ictalurus punctatus) neutrophil collagenase

Channel catfish (Ictalurus punctatus) neutrophils, like mammalian neutrophils, contain a variety of enzymes and lytic peptides that participate in pathogen destruction. We have identified and characterized from a channel catfish anterior kidney cDNA library a 1.6 kb cDNA that encodes for channel catfish neutrophil collagenase. The deduced amino acid sequence has a predicted molecular mass of 53 kDa. The putative catfish collagenase has nucleotide and amino acid homology of 51.4% and 45.1%, respectively, with human neutrophil collagenase and 50.4% and 47.1%, respectively, with mouse neutrophil collagenase. Certain regions of the molecule, including the cysteine switch and the putative zinc binding sites, were identical to those in the human and mouse genes. Polyclonal antiserum, prepared to the fusion protein, recognizes proteins from channel catfish neutrophil supernatants with molecular masses of approximately 63, 53 and 28 kDa. Supernatants from phorbol dibutyrate stimulated neutrophils were capable of degrading type I collagen. In addition, the polyclonal antiserum prevented the collagenase activity of the supernatants from stimulated catfish neutrophils; whereas, preimmune serum had no effect on collagenase activity of supernatants. Supernatants from unstimulated cells or the fusion protein did not possess the ability of degrading type I collagen. These results indicate that channel catfish neutrophil collagenases share molecular and functional features with mammalian neutrophil collagenase.     

Influence of route of administration on the humoral response of channel catfish (Ictalurus punctatus) to Yersinia ruckeri

Channel catfish were inoculated intraperitoneally, intramuscularly, or intraesophageally with Yersinia ruckeri. Three antigen doses were administered by each route of injection. Four fish from each treatment were sacrificed at 5-day intervals for 40 days. Serum from each individual was tested for antibody activity against Y. ruckeri by indirect enzyme-linked immunosorbent assay (ELISA). High titers of anti-Y. ruckeri antibody were elicited by all doses (10(5) to 10(9) cells/g of fish) regardless of the route of administration. Mean titers for saline injected fish ranged up to 1:64 for each route of inoculation while mean titers for bacteria injected fish ranged up to 1:4096. Mean titers of 1:128 or greater were observed by day 15 post injection; titers peaked about day 30 and diminished thereafter. Channel catfish were most responsive to the antigen (10(7) to 10(9) cells/g of fish) when administered intramuscularly although lower doses (10(5) to 10(6) cells/g of fish) administered intraperitoneally elicited a substantial response. There was little evidence of dose-dependent responses for any of the routes of immunization. The rapid onset of relatively high serum titers suggests that the fish were mounting a secondary response to Yersinia ruckeri.     

Plasma/muscle ratios of sulfadimethoxine residues in channel catfish (Ictalurus punctatus)

Channel catfish ( n = 84) maintained at a water temperature of 27°C were used in a feeding study to determine the plasma to muscle concentration ratios of sulfadimethoxine (SDM) and 4-N-acetylsulfadimethoxine residues. Sulfadimethoxine medicated feed was provided free choice at 42 mg SDM/kg body weight once daily for 5 days and the plasma and muscle concentrations of SDM were determined at selected withdrawal times (6, 12, 24, 48, 72, and 96 hours) following the last dose. Considerable variation in total SDM tissue concentration among fish within a sampling period was observed. For fish ( n = 12) at six hours post-dose, total SDM concentrations ranged from 1.4–24.8 μg/mL and 0.6–12.6 μg/g, with mean total SDM concentrations of 9.1 μg/mL and 5.3 μg/g for plasma and muscle, respectively. However, a mean plasma:muscle concentration ratio of 1.8:1 ± 0.3:1 was obtained over all concentrations and sampling periods. The plasma:muscle 95% t distribution interval for individual fish was 1.2:1 to 2.4:1. A correlation coefficient of 0.967 was obtained for the relationship between plasma and muscle total SDM concentration among individual fish ( n = 25). Results of this study indicate that plasma total SDM concentration may be used to identify samples containing violative SDM muscle residue. No fish contained total SDM muscle residues greater than the FDA tolerance (0.1 μg/g) by 48 hours following the final dose.     

Reciprocal translocation (13;20)(q12;q22) in an Icelandic sheep

Cytogenetic examination of G-banded lymphocyte chromosomes of an Icelandic ram from a line with a history of poor fertility revealed a rcp (13;20) (q12;q22) translocation. Meiotic studies showed a quadrivalent configuration at diakinesis and this was confirmed by C-banding.     

Reproductive consequences of an X-autosome translocation in a swine herd.

A survey was conducted on the chromosome makeup and breeding data of pigs housed at the Arkell Research Station, University of Guelph, to determine the reproductive impact of a reciprocal translocation previously identified as t(Xp+;14q-). Blood samples were cultured and chromosome spreads were examined to identify carriers in the herd. Gonadal features of carrier boars and piglets and their age-matched normal counterparts were studied, and the reproductive history of carrier sows, including the number of breedings required for conception, mean litter size, prevalence of still births and malformations, and sex distribution among the piglets, was compared with that of normal (noncarrier) sows. Our observations on testicular histology confirmed that X-autosome translocation-carrier boars are sterile, and the analysis of reproductive data showed that the most striking effects of t(Xp+;14q-) on sows are the 50% increase in the number of breedings required for conception and the over 25% reduction in litter size compared with that of normal sows. The reproductive impact of this and similar translocations on the herd and on the swine industry in general is discussed.     

Use of an aiming device for application of a type-II (bilateral) external fixator to a fractured tibia of a dog

An aiming device was used to guide insertion of fixation pins in a type-II (bilateral) external fixator stabilizing an open canine tibial fracture. This device, designed by the Swiss AO group, has multiple applications in orthopedic surgery, because it accurately locates the exit point of a pin or drill hole on the far side of a bone or fractured bone fragment. When used with the type-II external fixator, it greatly facilitates pin placement by ensuring that, as a pin emerges from the bone, it is in line with the second clamp on the opposite connecting bar.     

Development of a fast agglutination screening test (FAST) for the detection of anti-Leishmania antibodies in dogs

A fast agglutination screening test (FAST) for the detection of anti-Leishmania antibodies in serum samples from dogs with visceral leishmaniosis was developed. The test is based on the direct agglutination test (DAT), but combines a higher parasite concentration with a smaller test volume. In contrast to the DAT, the FAST makes use of only one serum dilution and the results can be read within 3 h as opposed to 18-20 h for the DAT. The FAST was evaluated using serum samples of confirmed cases of the disease and healthy controls collected in the most important endemic regions of canine visceral leishmaniosis, import cases of canine leishmaniosis in a non-endemic country, from non-endemic healthy controls and from dogs with other diseases. The performance of the FAST was compared with standard DAT. In the present study, the FAST had a sensitivity of 93.6% and a specificity of 89.0%. The DAT had a sensitivity of 88.6% and a specificity of 96.7%. Furthermore, using a large panel of serum samples of previously examined DAT positive or negative dogs it was shown that degree of agreement between the two tests was high (95.7%; kappa value = 0.91). The FAST offers the advantages of the DAT based on freeze-dried antigen with respect to stability of the antigen, sensitivity and specificity. Moreover, the FAST allows the rapid screening of a large number of samples, which makes the test very useful for epidemiological screening of large populations of dogs.     

植酸酶预处理饲料对斑点叉尾鮰生长及磷利用率的影响

本试验通过体外预处理方式在饲料中添加植酸酶,旨在探讨其对斑点叉尾鮰生长及磷利用率的影响.试验以初始条件一致、平均体重为(1.74±0.02)g的450尾斑点叉尾鮰(Icralurus punctatus)稚鱼为研究对象,采用单因素完全分组设计,随机分为5个组,每组3个重复,每个重复30尾鱼.D0组为对照组,饲喂基础饲料...     

A light and electron microscopic study of the leukocytes of channel catfish (Ictalurus punctatus)

A light and electron microscopic study of channel catfish (Ictalurus punctatus) white blood cells was carried out. The white blood cells of channel catfish consist of lymphocytes, neutrophils, thrombocytes and monocytes. A fifth cell type is illustrated and described which could not be characterized. These leukocytes are characterized and compared with their mammalian counterparts.     

Application of an enzyme-immunoassay (EIA) for rapid screening of 5 alpha-pregnane-3,20-dione (DHP) in blood plasma of the Asian elephant, Elephas maximus.

Populations of African (Loxodonta Africana) and Asian Elephants (Elephas maximus) in zoos and safari parks are at risk due to their low reproductive success. To extend the limited knowledge of their reproductive physiology, the development of easy and practical methods for the analysis of the relevant reproductive hormones is essential to support e.g. assisted reproduction. For the measurement of 5 alpha-pregnane-3,20-dione (DHP), the predominant ovarian gestagen in both species, an enzyme-immunoassay (EIA) based on commercial reagents was applied. Advantages of this EIA are the small volume of plasma needed for evaluation (5 microliters) and the possibility of direct processing without an extraction stage. The lower limit of detection was 0.16 ng/ml, the mean recovery was 101% and the mean coefficients of variation were 7.3% (intra-assay) and 9.9% (inter-assay). In Asian elephants, DHP levels reached 15 ng/ml during the luteal phase and up to 21 ng/ml during pregnancy. Estrous cycle lengths based on the lowest DHP concentrations varied from 12 to 20 weeks (mean 15.4 +/- 2.3). In two Asian elephant cows a calf was still-borne. Thereafter, the animals reassumed ovarian activity after approximately 8 and 13 weeks, respectively. In one animal estradiol implants for hormonal contraception caused a down regulation of the ovarian function as demonstrated by an irregular pattern of DHP secretion over a period of 48 weeks. We propose the direct DHP-EIA as a suitable method for reproductive monitoring in elephants, as it can be easily established in laboratories.     

Single intravenous and oral dose pharmacokinetics of florfenicol in the channel catfish (Ictalurus punctatus)

Plasma distribution and elimination of florfenicol in channel catfish were investigated after a single dose (10 mg/kg) of intravenous (i.v.) or oral administration in freshwater at a mean water temperature of 25.4 °C. Florfenicol concentrations in plasma were analyzed by means of liquid chromatography with MS/MS detection. After i.v. florfenicol injection, the terminal half-life (t(1/2)), volume of distribution at steady state (V(ss)), and central volume of distribution (V(c)) were 8.25 h, 0.9 and 0.381 L/kg, respectively. After oral administration of florfenicol, the terminal t(1/2), C(max), T(max), and oral bioavailability (F) were 9.11 h, 7.6 μg/mL, 9.2 h, and 1.09, respectively. There was a lag absorption time of 1.67 h in oral dosing. Results from these studies support that 10 mg florfenicol/kg body weight in channel catfish is an efficacious dosage following oral administration.     

Sphaerospora infection of American catfishes (Ictalurus punctatus and I. nebulosus) in Europe.

The occurrence in Europe of a Sphaerospora species described in North America is reported. Based upon its morphological characteristics, the parasite could be identified with the species S. hankai described from brown bullhead in Canada. This parasite was found to infect channel catfish (Ictalurus punctatus) cultured in farm ponds in Italy and brown bullhead (Ictalurus nebulosus) living in the supply channels of fish ponds in Hungary. The spores and sporogonic developmental stages were situated in the lumen of the renal tubules. In the authors' opinion, S. ictaluri described from channel catfish can be considered synonymous with S. hankai.