Dermatomycosis caused by Paranannizziopsis australasiensis in five tuatara (Sphenodon punctatus) and a coastal bearded dragon (Pogona barbata) in a zoological collection in New Zealand

CASE HISTORY: Health monitoring of tuatara (Sphenodon punctatus) at Auckland Zoo between 2001 and 2009 showed that 58/93 tuatara had been affected by dermatitis of unknown origin. From 2011 onwards, cases of suspected fungal dermatitis underwent extensive diagnostic investigations.

CLINCAL FINDINGS: Six cases of dermatomycosis were attributed to Paranannizziopsis australasiensis, five in tuatara and one in a coastal bearded dragon (Pogona barbata). Cases presented typically as raised, yellow to brown encrustations on the skin. Severe cases progressed to necrotising ulcerative dermatitis, and in the bearded dragon to fatal systemic mycosis. Following topical and systemic treatments, lesions resolved in all five tuatara.

LABORATORY FINDINGS: Histopathological examination of skin biopsy samples revealed dermatitis with intralesional septate branching hyphae. Fungal culture yielded isolates morphologically resembling Chrysosporium species, and isolates were submitted for molecular confirmation and sequencing of DNA.

DIAGNOSIS: All six cases were confirmed as dermatitis due to infection with P. australasiensis, on the basis of fungal culture and DNA sequencing of isolates.

CLINICAL RELEVANCE: These are the first reported cases of dermatomycosis associated with P. australasiensis infection in tuatara, and the first cases in which systemic therapeutic agents have been used in the treatment of such disease. Tuatara at the Auckland Zoo are now routinely examined every 3 months and tissue samples from any lesions sent for histopathology and fungal culture. Further work to elucidate the epidemiology and significance of P. australasiensis infections in reptiles in New Zealand is important for both welfare and conservation purposes.     

Identification of duck HSP70 gene,polymorphism analysis and tissue expression under control and heat stress conditions

1. This study was designed to characterise the duck heat shock protein 70 gene (HSP70) and identify sequence variation.

2. Chicken HSP70 sequence (GenBank: AY143693) was used as a template to design a primer pair to amplify partial duck HSP70 gene. Primers were subsequently designed with the duck HSP70 gene as a template to amplify the complete duck HSP70 sequence.

3. Twelve commercial Sanshui White ducklings were subjected to a heat stress experiment. Tissue samples were collected for RNA extraction and real-time PCR to analyse the expression mechanism of duck HSP70.

4. A DNA pool was constructed from three different species for single nucleotide polymorphism (SNP) screening. The genotypes of the identified SNPs were detected in 145 Sanshui White ducklings.

5. Duck HSP70 gene was identified and characterised (GenBank: EU678246) and shown to contain no introns. Fifteen variations were identified within the open reading frame. Quantitative real-time PCR results showed that the expression of duck HSP70 gene was tissue specific and the highest expression level was seen in pectoral muscle.     

Gene expression profile of IGF1 and MSTN mRNA and their correlation with carcass traits in different breeds of geese at 70 d of age

1. The expression of insulin-like growth factor-1 (IGF1) and myostatin (MSTN) mRNA in breast and leg muscle was quantified in 70-d-old Taihu and Wanxi geese by using a Multiplex Competitive Fluorescent–PCR method and the correlations between mRNA levels and carcass traits were analysed.

2. IGF1 mRNA expression in breast muscle in Taihu geese was significantly higher than that in Wanxi geese and the MSTN mRNA level in leg muscle in Taihu geese was significantly higher than that in Wanxi geese.

3. There was no significant difference in breast muscle MSTN or leg muscle IGF1 mRNA expression between the two breeds.

4. Within the same breed, the IGF1 mRNA expression in leg muscle of male geese was significantly higher than that in female geese, and MSTN mRNA expression in leg muscle was significantly higher than that in breast muscle.

5. There was no difference in the IGF1 mRNA expression between tissues.

6. There was a positive correlation between IGF1 mRNA and MSTN mRNA and a negative correlation between IGF1 mRNA expression of breast muscle and leg muscle ratio.

7. In Wanxi geese, MSTN mRNA expression in leg muscle was negatively associated with body weight and leg muscle weight.     

Cellular changes in the granulosa layer of the maturing ovarian follicle of the domestic fowl

1. The relationships between length, width and height of the maturing ovarian follicle of the domestic fowl (Gallus domesticus) are shown.

2. During the rapid‐growth phase (yolk deposition) of the oocyte, i.e. an increase in size from about 9 mm diameter to 35 mm diameter, there is about a 15‐fold increase in its surface area. During this time the number of granulosa cells adjacent to the oocyte increases by about 5‐fold.

3. The granulosa cells become flatter during the growth period of the follicle, consequently increasing their surface area adjacent to the oocyte by about 3‐fold.

4. Together the change in shape of the granulosa cells and the increase in their number account for the increase in area of the granulosa layer during follicular growth.

5. Measurement of the DNA content of the granulosa layer indicated a progressive decrease in the cellular content of DNA as the follicle matures.     

Genotypic and phenotypic diversity differences of presumptive commensal and avian pathogenic E. coli

1. The objective of the experiment was to characterise the genotypic and phenotypic differences between presumptive commensal E. coli and avian pathogenic E. coli (APEC) of poultry.

2. DNA was extracted from 65 confirmed APEC E. coli from chicken, 100 presumptive commensal E. coli from healthy turkey and 35 from healthy chicken. Enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and virulence factors genotyping was performed to characterise genetic features.

3. Carbon source utilisation and antimicrobial susceptibility tests were performed to characterise phenotypic features of isolates.

4. The genetic divergence between E. coli strains tested by ERIC-PCR profiles and virulence-associated genes showed a clear genetic separation between E. coli APEC and turkey E. coli strains.

5. The carbon utilisation profile of turkey isolates was different from chicken and APEC strains; whereas antimicrobial susceptibility was highest for turkey isolates (53%), and lowest for APEC strains (33.8%).

6. The study showed a significant negative correlation between utilisation of arabitol and adonitol with different virulence determinants tested, which suggests that the ability to utilise some uncommon carbon sources may be used to discriminate between presumptive commensal E. coli and APEC.     

Effect of dietary supplementation of bacteriophage on performance,egg quality and caecal bacterial populations in laying hens

1. Bacteriophages (BP) have gained increasing attention as a treatment of bacterial infection for animals. However, the data pertaining to dietary application of BP for laying hens have been limited.

2. This study aimed to investigate the effect of dietary BP on laying performance, egg quality and caecal bacterial populations in laying hens.

3. The dietary BP used in this experiment was a mixture of individual BP targeting Salmonella gallinarum, Salmonella pullorum, Salmonella typhimurium, Salmonella enteritidis, Salmonella derby and Staphylococcus aureus.

4. A total of 360 Hy-Line Brown laying hens of 32 weeks of age were allotted to one of three dietary treatments with 6 replicates in a completely randomised design. The basal diet was prepared, and 0.4 or 0.8 g/kg BP mixture was supplemented to the basal diet. Diets were fed to hens for 8 weeks.

5. Laying performance and egg quality were not affected by dietary treatments. As inclusion levels of BP mixture in diets were increased, the DNA copy numbers for Salmonella spp. in the caecal contents decreased linearly, whereas the DNA copy numbers for Escherichia coli in the caecal contents increased linearly.

6. Results indicate that dietary supplementation of BP mixture decreases the target Salmonella spp. populations but increases Escherichia coli populations in the gastrointestinal tract of laying hens with little impact on laying performance and egg quality.     

Molecular typing of Iranian field isolates Mycoplasma synoviae and their differentiation from the live commercial vaccine strain MS-H using vlhA gene

1. The single-copy domain of the N-terminal region of the vlhA gene of Mycoplasma synoviae was sequenced, analysed and verified and used to type Iranian field isolates of M. synoviae and the MS-H live vaccine strain. In addition, a restriction fragment length polymorphism (RFLP) method was developed to differentiate between field isolates of Iranian and MS-H vaccine strains.

2. All sequences were analysed and aligned; the percentage similarity of the DNA was calculated and dendrograms were constructed. Based on single nucleotide polymorphism (SNP) that existed in all field isolates in Iran, the PCR-RFLP method allowed the differentiation of all M. synoviae field isolates from the vaccine strain.

3. Using phylogenetic analysis, the isolates were assigned to 8 unique genotypes and, within each group, DNA had a high level of similarity.

4. DNA sequence analysis and PCR-RFLP of the amplicon based on percent similarity and evolutionary relationship appeared to be useful tools for strain differentiation whether M. synoviae clinical isolates from infected chickens were derived from the vaccine strain or wild-type strains.

5. This study confirms the potential value of strain typing for epidemiological purposes and suggests that phylogenetic studies are essential to understand the true relationships between strains.     

Paired box 7 (Pax7) gene: molecular characterisation,polymorphism and its association with growth performance in goose (Anser cygnoides)

1. Paired box (Pax7) gene is a member of the paired box family and plays a critical role in animal growth and muscle development. However, the molecular characterisation of the goose Pax7 gene is unknown.

2. The open-reading frame of goose Pax7 is composed of 1509 bp, which encodes a protein of 503 amino acids and shares high homology with Pax7 of other birds.

3. Ten single-nucleotide polymorphisms were identified in the genomic DNA sequence, 8 located in the intron region and two located in the exon region.

4. Association analysis showed the C122T locus was significantly associated with the body weight of Zhedong-White geese in week 4, 6, 8, 10 and 12.

5. It was concluded that the goose Pax7 gene may be an important candidate gene for goose growth traits and marker-assisted selection.     

A mathematical representation of the Ovulatory cycle of the domestic hen

1. The ovulatory cycle of the domestic hen (Gallus domesticus) was postulated to be the result of the interaction of two independent systems.

2. A circadian system was postulated to control the restriction of ovulation to an 8‐h period of the day under conventional 14 h light: 10 h dark regimes.

3. The final phase of follicular maturation was postulated to commence after ovulation of the preceding ovum in the hierarchy.

4. Ovulation was postulated to occur when a mature follicle was present in the ovary during the appropriate phase of the circadian‐linked system.

5. The predicted times of oviposition were within the standard error of the observed times of oviposition under 21‐, 24‐ and 28‐h photoschedules.

6. It was concluded that this hypothesis for the control of the ovulatory cycle of the hen is consistent with current knowledge.     

The effects of restricted feeding on adrenal cortical activity in the immature domestic fowl

1. The effect of reducing food intake to 75% of the ad libitum intake was determined from hatching to 8 weeks in young Light Sussex chickens.

2. Restricted birds were lighter throughout the experiment.

3. Relative adrenal weight tended to be greater in restricted birds but the difference decreased with time.

4. There was no depletion of adrenal cholesterol: from week 5 there was a significantly greater amount in the adrenals of restricted birds.

5. After 1 week of restriction plasma corticosterone concentration was 73% greater than in controls. It decreased progressively, falling within the normal range at 5 weeks.

6. Restricted birds were hypoglycaemic from weeks 2 to 7 and hyper‐lipacidaemic throughout. A negative correlation between plasma glucose and free fatty acids was found.     

Encapsulated benzoic acid supplementation in broiler diets influences gut bacterial composition and activity

1. The present study investigated the effects of encapsulated benzoic acid (BA) supplementation in broiler feed on performance and gastrointestinal microbiota.

2. Eighty broilers were randomly divided into two groups. Birds in the control group were fed on maize–soybean-based diets. Birds in the treatment group were provided the same diet supplemented with 2 g/kg BA encapsulated in a vegetable oil matrix.

3. At the end of the trial (d 35), pH, bacterial composition and metabolites were determined in the crop, jejunum, ileum and caecum.

4. Growth performance variables and pH were not significantly different.

5. BA concentration decreased rapidly in the proximal gut. However, the treatment diet showed higher BA in the crop, jejunum, ileum and caecum.

6. Total lactate in the crop and D-lactate in the jejunum was higher in the BA treated group. Caecal total and branched chain fatty acids were decreased due to the treatment.

7. Lactobacilli populations were significantly altered by BA supplementation. A trend for increased lactobacilli was observed in the crop, while it became significant in the jejunum and ileum. Lactobacillus species responded differently to the treatment. Four of 5 measured Lactobacillus species, particularly in the ileum, followed the course observed for total lactobacilli; only Lactobacillus salivarius was not modified.

8. Correlation analysis showed that BA modified the intestinal microbiota. Lactobacilli correlated negatively to all studied clostridial clusters and enterobacteria. Clostridial clusters IV and XIVa were significantly increased in the jejunum, whereas only clostridial cluster XIVa was increased in the caecum.

9. Encapsulated BA modified the intestinal microbiota which can lead to the conclusion, that the main beneficial mode of action of BA in the gut appears to be the enhancement of lactic acid bacteria, which in turn may act as a vanguard against pathogens.     

Effects of dietary supplementation with Gynura procumbens (Merr.) on egg yolk cholesterol,excreta microflora and laying hen performance

1. The present study investigated the effects of dietary supplementation with Gynura procumbens on egg yolk and serum cholesterol and triglycerides, excreta microflora, laying performance and egg quality.

2. A total of 160 Hy-Line Brown layers (45 weeks old) were randomly assigned into 4 treatments on the basis of laying performance. Each treatment had 4 replicates with 10 birds each.

3. Dietary treatments were basal diet supplemented with 0 (control), 2.5, 5.0 and 7.5 g/kg diet G. procumbens during 56-d feeding period.

4. Serum (d 21, 42 and 56) and egg yolk (d 28, 42 and 56) cholesterol and triglycerides concentrations were linearly reduced with increasing dietary concentrations of G. procumbens.

5. Increasing dietary concentrations of G. procumbens linearly reduced the excreta total anaerobic bacteria (d 28), Clostridium sp. and Escherichia coli (d 28 and 56) populations.

6. Overall egg production and egg mass were linearly increased, and overall feed efficiency was linearly improved with increase in dietary G. procumbens.

7. Dietary increasing concentrations of G. procumbens linearly improved egg yolk colour (d 28 and 56) and breaking strength of eggs (d 56).

8. The results obtained in the present experiment indicate that dietary supplementation with G. procumbens could reduce the egg yolk cholesterol, suppresses harmful excreta microflora and improves layers performance.     

Isolation and molecular characterisation of chicken parvovirus from Brazilian flocks with enteric disorders

1. The presence of parvovirus in chickens with enteric disease was investigated in commercial flocks in Brazil.

2. The intestinal contents of chickens exhibiting clinical signs of diarrhoea, weight loss or mortality were examined, and chicken parvovirus (chPV) was identified using a polymerase chain reaction (PCR) assay. The samples were sequenced and inoculated into specific-pathogen-free (SPF) embryonated eggs to isolate the virus.

3. Necropsies showed that the embryos were dwarfish, haemorrhagic and oedematous. The presence of chPV was confirmed by PCR and DNA sequencing.

4. The molecular characterisation of chPV strains circulating in the Brazilian flocks showed that they were genetically related to sequences from North America, Europe and Asia. Phylogenetic analyses clustered the Brazilian chPV sequences with those from Europe (Croatia, Hungary) and Asia (South Korea).

5. This study is the first report of the molecular characterisation of chPV circulating in the commercial flocks in Brazil and indicates high genetic similarity with chPV sequences from around the world.     

The occurrence of giant fibres in different muscles of two chicken genotypes

1. The occurrence of Giant Fibres (GF) in three muscles (Pectoralis major (PM), Iliotibialis lateralis and Semimembranosus) with different types of energy metabolism was studied in slow- and fast-growing chicken strains.

2. A total of 20 one-day-old Leghorn chicks (slow-growing) and 20 broiler (Ross 508) chicks (fast-growing) were reared to 100 and 45 d, respectively.

3. A small percentage of GF was seen in pre rigor muscle samples even at 3 min post mortem in both genotypes and in all muscle types studied.

4. From 3 min to 24 h post mortem GF increased both in Leghorn and broiler chickens but to a different extent according to muscle type and genotype.

5. The highest GF 24 h post mortem value was found in the PM muscles belonging to the fast-growing broiler line.

6. It was concluded that every type of muscle can develop GF, but this phenomenon is more evident in the PM especially in animals selected for increased growth rate.     

Investigation of cell-free DNA in canine plasma and its relation to disease

Background: DNA is released from dying cells during apoptosis and necrosis. This cell-free DNA (cfDNA) diffuses into the plasma where it can be measured. In humans, an increase in cfDNA correlates with disease severity and prognosis.

Objective: It was hypothesized that when DNA in canine plasma was measured by emission fluorometry without prior DNA extraction, the concentration of cfDNA would increase with disease severity.

Animals: The diseased population consisted of 97 client-owned dogs. The clinically normal population consisted of nine client-owned dogs presenting for ‘wellness screens’, and 15 colony-owned Harrier Hounds.

Methods: Plasma cfDNA was measured by fluorometry without prior DNA extraction. The effects of ex vivo storage conditions were evaluated in plasma from two clinically normal dogs. In all other dogs, plasma was separated within two hours of collection. The association between the cfDNA concentration in hospitalized dogs and a variety of clinical, clinicopathological and outcome variables was tested.

Results: The concentration of cfDNA was reliably measured when plasma was separated within two hours of blood collection. The diseased dogs had significantly higher cfDNA than clinically normal dogs (P < 0.001), and the more severe the disease, the higher the cfDNA when severity was categorized according to the American Society of Anesthesiologists (ASA) status (P < 0.001). Dogs that did not survive to discharge had significantly higher cfDNA concentrations than survivors (P = 0.02).

Conclusions/Clinical Importance: The concentration of cfDNA in the plasma of diseased dogs is associated with disease severity and prognosis. Measurement of canine cfDNA could be a useful non-specific disease indicator and prognostic tool.     

Molecular serotyping and antimicrobial resistance profiles of Actinobacillus pleuropneumoniae isolated from pigs in South Korea

Background: Actinobacillus pleuropneumoniae (APP) causes porcine pleuropneumonia (PP).

Objective: Serotypes and antimicrobial resistance patterns in APP isolates from pigs in Korea were examined.

Methods: Sixty-five APP isolates were genetically serotyped using standard and multiplex PCR (polymerase chain reaction). Antimicrobial susceptibilities were tested using the standardized disk-agar method. PCR was used to detect β-lactam, gentamicin and tetracycline-resistance genes. The random amplified polymorphic DNA (RAPD) patterns were determined by PCR.

Results: Korean pigs predominantly carried APP serotypes 1 and 5. Among 65 isolates, one isolate was sensitive to all 12 antimicrobials tested in this study. Sixty-two isolates was resistant to tetracycline and 53 isolates carried one or five genes including tet(B), tet(A), tet(H), tet(M)/tet(O), tet(C), tet(G) and/or tet(L)-1 markers. Among 64 strains, 9% and 26.6% were resistance to 10 and three or more antimicrobials, respectively. Thirteen different antimicrobial resistance patterns were observed and RAPD analysis revealed a separation of the isolates into two clusters: cluster II (6 strains resistant to 10 antimicrobials) and cluster I (the other 59 strains).

Conclusion: Results show that APP serotypes 1 and 5 are the most common in Korea, and multi-drug resistant strains are prevalent. RAPD analysis demonstrated that six isolates resistant to 10 antimicrobials belonged to the same cluster.     

A 31P topical magnetic resonance study of embryonic development in hens' eggs

1. A ³¹P topical magnetic resonance study of whole chicken's eggs in vivo is described.

2. Resonances from adenosine triphosphate, phosphocreatine, inorganic phosphate and phosphorous storage protein were observed. These changed in relative intensity as the embryo developed.

3. No ill effects due to exposure to radiofrequency radiation or magnetic fields were observed.

4. It is concluded that topical magnetic resonance can make a useful contribution to the study of embryonic development in avian eggs.     

Comparison of the productive perform. Ances of the Sinai Bedouin fowl,the white leghorn and their crossbreds: Study under laboratory conditions

1. A comparison of the productive performances of the Sinai Bedouin fowl (from the Sinai desert), the commercial White Leghorn and their reciprocal crossbreds was made under uniform normal conditions.

2. The Sinai produced significantly fewer, smaller eggs, resulting in lower egg mass output (g/bird d), than the Leghorn and the crossbreds.

3. Egg weight of both crossbreds was intermediate between Sinai and Leghorn but laying rate was closer to that of the Leghorn.

4. Differences in egg mass output apparent at 8 to 10 months of age decreased considerably with age.

5. These findings suggest that selection or crossbreeding of the Sinai might improve productive performance while maintaining improved eggshell quality.     

Occurrence of Arcobacter in Iranian poultry and slaughterhouse samples implicates contamination by processing equipment and procedures

1. The occurrence of Arcobacter spp. and three pathogenic species of Arcobacter from Iranian poultry carcasses was investigated at different steps of broiler processing to determine critical control points for reducing carcass contamination.

2. Samples were collected from (a) cloaca immediately before processing, (b) different points during processing and (c) at different stations in a processing plant of a slaughterhouse in southern Iran.

3. After enrichment steps in Arcobacter selective broth, DNA of the samples was extracted and three significant pathogen species of Arcobacter were identified based on polymerase chain reaction (PCR) detection of 16S rRNA and specific species PCR.

4. Out of a total of 540 samples, 244 (45%) were positive for Arcobacter spp. Arcobacter butzleri was more frequently detected (73% ± 13.9%) than A. cryaeophilus (9% ± 13.9%) and A. skirrowii (4.1%). In addition, co-colonisation (A. butzleri and A. cryaerophilus) occurred in 13.9% of the positive samples.

5. The results indicate a high prevalence of Arcobacter in the investigated slaughterhouse and broiler carcasses and that Arcobacter is not a normal flora of the broilers. Evidence for the presence of Arcobacter in the environment and water of processing plants suggests that these are sources of contamination of poultry carcasses. In addition, contamination of the poultry carcasses can spread between poultry meats in different parts and processes of the slaughterhouse (pre-scalding to after evisceration).     

Influence of ascorbic acid on the adverse effect of feeding a diet containing excess tyrosine to chicks

1. The effects of dietary or parenteral administration of ascorbic acid on the adverse effects of excess dietary tyrosine were investigated with young male White Leghorn chicks in a 2‐week experiment.

2. Addition of 10 g ascorbic acid/kg to the control diet (without excess tyrosine) produced no beneficial effects on performance. Excess dietary tyrosine caused depressions in all measures of performance.

3. Adding 0.1, 1, 10 or 20 g ascorbic acid/kg to the diet containing excess tyrosine tended to improve performance.

4. Subcutaneous injection of 50 mg ascorbic acid/bird d to chicks receiving excess tyrosine brought about a significant improvement in body‐weight gain.

5. The elevation of plasma free tyrosine caused by excess dietary tyrosine decreased as dietary ascorbic acid increased.

6. Ascorbic acid can alleviate, though not completely counteract, the adverse effect of excess dietary tyrosine.