Molecular sexing of Jungle crow (Corvus macrorhynchos japonensis) and Carrion crow (Corvus corone corone) using a feather

A molecular sexing method using the intron‐size difference between chromo‐helicase‐DNA binding protein (CHD1) genes on the Z and W sex chromosomes (CHD1Z and CHD1W, respectively) was developed for wild Jungle crows and wild Carrion crows. The polymerase chain reaction product sizes were 661 bp for CHD1Z and 465 bp for CHD1W in both the Jungle and the Carrion crows. Each male crow sample produced a single fragment (CHD1Z), whereas female samples produced two fragments (CHD1Z and CHD1W), which differed by 200 bp in crows. In the present study, sexing live Jungle and Carrion crows was performed by applying the method to a feather for molecular sexing.     

West Nile virus antibody prevalence in American crows (Corvus brachyrhynchos) and fish crows (Corvus ossifragus) in Georgia, USA

Crows have been the centerpiece of avian West Nile virus (WNV) surveillance and research in North America. This work has demonstrated variation in susceptibility to WNV infection between American (Cor vus brachyrhynchos) andFish Crows (Corvus ossifragus). The higher WNV-associated mortality rate in American Crows compared with Fish Crows suggests that WNV antibody prevalence would be greater in the Fish Crow population. The objectives of this study were to 1) determine whether Fish Crows had higher WNV antibody prevalencethan American Crows, 2 ) determine th e persistence o f antibodies to WNV in naturally infected Fish Crows, and 3) develop a technique to distinguish Fish Crows from American Crows on the basis of sequence analysis and restriction enzyme digestion of a mitochondrial DNA fragment. West Nile virus antibody prevalence was 16.5% (n = 97) in Fish Crows and 5.7% in American Crows (n = 53) collected from Georgia between 2004 and 2006. Antibodies persisted at high titers for 12 mo in Fish Crows. This is the first report of WNV antibody persistence in a crow species. A polymerase chain reaction technique paired with restriction enzyme digestion easily distinguished American Crows from Fish Crows on the basis of a mitochondrial DNA fragment.     

Prevalence of thermophilic campylobacter infections in humans, chickens and crows in Morogoro, Tanzania

Prevalence of thermophilic Campylobacter infections in humans, chickens and crows was determined in a cross-sectional study that was carried out in urban and rural areas of Morogoro region, Tanzania during the period of January 2003 to December 2004. A total of 632 human stool samples, 536 cloacal swabs from local and broiler chickens and 22 intestinal contents from crows were screened for presence of thermophilic campylobacters using Skirrow's protocol. Representative Campylobacter jejuni isolates from human and chicken samples were also analysed by polymerase chain reaction (PCR) as a definitive identification method. The overall prevalence of thermophilic campylobacters was 9.3% (95% CI: 7.2-11.9), 69.8% (95% CI: 65.7-73.6) and 72.7% (95% CI: 49.8-89.3) in humans, chickens and crows respectively. In humans, 59 thermophilic campylobacters were isolated of which 96.6% were C. jejuni and 3.4%Campylobacter coli. There was a significantly (P<0.001) higher prevalence in young individuals (16%) than in adults (7%). Of 341 isolates from chickens, 91.2% were C. jejuni and 8.8% were C. coli. A significantly (P<0.05) higher infection rate was observed in rural local chicken (76%) than in broilers (60%). In crows, of 16 isolates, 93.8% were C. jejuni and 6.2% were C. coli. Definitive identification of C. jejuni by PCR revealed positive results in 74.1% of 243 analysed isolates. Findings in this study indicate high prevalence of thermophilic campylobacters in humans, chickens and crows in Morogoro, and a higher infection rate of C. jejuni than that of C. coli in different animal species. Age of humans and location of chickens were identified as risk factors for thermophilic Campylobacter infections. Positive isolates to biochemical tests that indicated negative results on PCR indicates the additional value of PCR for definitive diagnosis of C. jejuni.     

Survey of campylobacter, salmonella and mycoplasmas in house crows (Corvus splendens) in Malaysia

House crows (Corvus splendens) in Selangor, Malaysia were examined for the presence of Campylobacter species, Salmonella species, Mycoplasma gallisepticum and Mycoplasma synoviae by serology, culture and pcr. For the detection of Campylobacter and Salmonella species swabs were taken either from the intestine or cloaca. For the detection of mycoplasmas, swabs were taken either from the choanal cleft or trachea for culture and pcr and serum samples were tested by the rapid serum agglutination (rsa) and monoclonal antibody-blocking elisa (mbelisa) for antibodies to M gallisepticum and M synoviae. For campylobacter, 25.3 per cent of the crows were positive by culture, and the species identified were Campylobacter jejuni and Campylobacter coli. No Salmonella species were isolated. Four of 24 swabs were positive for M gallisepticum dna but none gave positive results for M synoviae dna. No M gallisepticum or M synoviae antibodies were detected by rsa but 60 per cent of the sera gave positive reactions for M gallisepticum and 13 per cent gave positive reactions for M synoviae by mbelisa.     

Characterization of Campylobacter jejuni in large-billed crows (Corvus macrorhynchos) in Tochigi prefecture,Japan

As free-living crows are a potential source of Campylobacter infections in broilers and cattle, we characterized Campylobacter spp. isolated from crows using multilocus sequence typing and antimicrobial susceptibility testing. We obtained 82 samples from 27 birds captured at seven different times using a trap set in Tochigi prefecture, Japan. Campylobacter jejuni was isolated from 55 (67.1%) of the 82 samples and classified into 29 sequence types, of which 21 were novel. Tetracycline and streptomycin resistance rates were 18.2% and 3.6%, respectively. These results show that most types of C. jejuni infecting crows differ from those isolated from humans, broilers, and cattle. Thus, the importance of free-living crows as reservoirs of Campylobacter infections in broilers and cattle may be limited.     

Clinical and pathologic responses of American crows (Corvus brachyrhynchos) and fish crows (C ossifragus) to experimental West Nile virus infection

West Nile virus (WNV)-associated disease has a range of clinical manifestations among avian taxa, the reasons for which are not known. Species susceptibility varies within the avian family Corvidae, with estimated mortality rates ranging from 50 to 100%. We examined and compared virologic, immunologic, pathologic, and clinical responses in 2 corvid species, the American crow (Corvus brachyrhynchos) and the fish crow (C ossifragus), following experimental WNV inoculation. Unlike fish crows, which remained clinically normal throughout the study, American crows succumbed to WNV infection subsequent to dehydration, electrolyte and pH imbalances, and delayed or depressed humoral immune responses concurrent with marked, widespread virus replication. Viral titers were approximately 3,000 times greater in blood and 30,000 to 50,000 times greater in other tissues (eg, pancreas and small intestine) in American crows versus fish crows. Histologic lesion patterns and antigen deposition supported the differing clinical outcomes, with greater severity and distribution of lesions and WNV antigen in American crows. Both crow species had multiorgan necrosis and inflammation, although lesions were more frequent, severe, and widespread in American crows, in which the most commonly affected tissues were small intestine, spleen, and liver. American crows also had inflammation of vessels and nerves in multiple tissues, including heart, kidney, and the gastrointestinal tract. WNV antigen was most commonly observed within monocytes, macrophages, and other cells of the reticuloendothelial system of affected tissues. Collectively, the data support that WNV-infected American crows experience uncontrolled systemic infection leading to multiorgan failure and rapid death.     

Experimental inoculation of European starlings (Sturnus vulgaris) and American crows (Corvus brachyrhynchos) with Mycobacterium bovis

The purpose of this series of pilot studies was to determine whether the passerine species studied are susceptible to infection with Mycobacterium bovis. Separate experiments were conducted on wild-caught starlings (Sturnus vulgaris) and American crows (Corvus brachyrhynchos). In each experiment, four birds were challenged intraperitoneally and four were challenged orally with microorganisms. Challenge dose was 1 x 10(5) colony-forming units of M. bovis cultured from a white-tailed deer (Odocoileus virginianus) case in Michigan. Birds were euthanatized at 1 and 2 mo postinoculation. Histologic lesions suggestive of mycobacteriosis, without the presence of acid-fast bacilli, were noted in all experimental groups. Mycobacterial cultures performed on pooled tissue samples were positive for M. bovis in only some of the intraperitoneal inoculates of each species.     

Nutritional secondary hyperparathyroidism in free-living fledgling American crows (Corvus brachyrhynchos brachyrhynchos).

From June 1994 to June 1996, 18 fledgling American crows (Corvus brachyrhynchos brachyrhynchos) from multiple locations on Long Island, New York, were presented with signs of metabolic bone disease characterized by folding fractures of the proximal tibiotarsus. Plasma alkaline phosphatase levels were elevated, and the calcium/phosphorus ratio and 25-hydroxycholecalciferol (25-(OH)D3) levels were decreased. The histopathologic diagnosis was parathyroid hyperplasia and generalized osteodystrophia fibrosa. A diet low in bioavailable calcium and/or vitamin D3 was the probable cause. Also, exposure to xenobiotics could have contributed to the depression of 25-(OH)D3 levels.     

Prevalence and antibiotic susceptibility of thermophilic Campylobacter isolates from free range domestic duck (Cairina moschata) in Morogoro municipality, Tanzania

Prevalence and antibiotic susceptibility of thermophilic Campylobacter isolated from free-ranging ducks was determined in Morogoro Municipality, Tanzania. Ninety intestinal contents from ducks were screened for thermophilic Campylobacter using Skirrow’s protocol. Of the Campylobacter jejuni isolates, 50 were tested for sensitivity to 12 antibiotics. Overall prevalence of thermophilic Campylobacter was 80%. The prevalence of Campylobacter in adult ducks (91.3%) was significantly (p?<?0.05) higher than ducklings (68.2%). The isolation rate of C. jejuni (81.9%) was significantly (P?<?0.001) higher than C. coli (18.1%). All C. jejuni isolates were susceptible to streptomycin, nitrofurantoin and amikacin. Forty eight percent, 74% and 82% of isolates were resistant to cefuroxime sodium, tetracycline and ampicillin respectively. Between 20–50% of isolates were resistant to erythromycin, gentamicin, cloxacillin and amoxicillin. Norfloxacin and ciprofloxacin had lower C. jejuni resistance of 10% and 16% respectively. C. jejuni isolates from adult ducks showed significantly higher rates of resistance (p?<?0.05) to most antibiotics than did duckling isolates. High prevalence of thermophilic Campylobacter in ducks could be of public health significance in Morogoro municipality. The observed multidrug resistance in this study poses a threat of transfer of antibiotic resistance to human pathogens because of the close contact between ducks and human.     

Introduction and spread of thermophilic campylobacters in broiler flocks.

Campylobacteriosis is the most commonly reported infectious cause of human gastroenteritis in developed countries and broiler chickens are considered to be the major food-borne source of the infection. The control of the infection in man depends upon its control in broiler flocks but the epidemiology in poultry is poorly understood. Up to 50 per cent of broiler flocks may be infected and most of the birds in an infected flock carry the organisms until slaughter. Vertical transmission through the egg appears unlikely but there are many other potential sources of the infection for the chicks; direct contact with infected animals or birds has been proposed and there is also evidence for indirect transmission through drinking water or insect vectors. It is suggested that the cross-sectional studies discussed in this review should be followed by well designed case-control studies to test the aetiological hypotheses put forward.     

First molecular identification and subtype distribution of Blastocystis sp. isolated from hooded crows (Corvus cornix) and pigeons (Columba livia) in Tehran Province,Iran

Blastocystis is a common intestinal parasite among humans and animals such as non-human primates, pigs, cattle, birds, amphibians, and less frequently, rats, reptiles and insects. Since Blastocystis is a widely transmissible parasite between humans and mammals or birds, it is prominent to determine whether newly secluded non-human isolates are zoonotic. There are no comprehensive studies in Iran assessing the prevalence and molecular identification of Blastocystis infection in birds, especially in pigeons and crows. So, the aim of this study was to identify Blastocystis subtypes (STs) in crows and pigeons in Tehran province, Iran, using Nested PCR-RFLP and sequencing. Overall, 300 Blastocystis isolates from birds (156 pigeons and 144 crows) were subtyped by PCR, and the homology among isolates was then confirmed by RFLP analysis of the 18S rRNA gene. The prevalence of Blastocystis infection was detected 42.9% in pigeons and 44.4% in crows. All positive pigeons were owned by ST13 (100%). Among crows, 46 samples (71.8%) like pigeons were ST13, and 13 samples (20.3%) were ST14. Five samples (7.9%) remained unknown. This study was the first report of ST13 and ST14 of Blastocystis from birds. In the present study, our data revealed a high prevalence of Blastocystis sp. in pigeon’s and crow’s samples and the isolates from these birds were classified into two genetically distinct STs. Therefore, birds appear to be infected with various STs. It is important to determine the phylogenetic relationships between unknown STs from these birds and the multiple STs of Blastocystis.     

Prevalence of thermophilic Campylobacter species in Swedish dogs and characterization of C. jejuni isolates

Background

The aims of this study were to investigate the prevalence of Campylobacter species in Swedish dogs, to identify the species of the Campylobacter isolates and to genotype the C. jejuni isolates. Young and healthy dogs were targeted and the sampling was performed at 11 veterinary clinics throughout Sweden from October 2011 to October 2012. Faecal swab samples were collected and sent to the laboratory at the National Veterinary Institute (SVA) for isolation of Campylobacter, speciation and genotyping.

Results

Campylobacter spp. were isolated from 67 of the 180 sampled dogs which yields an overall prevalence of 37%. The most prevalent species of Campylobacter among the participating dogs was C. upsaliensis with 52 of the 67 identified isolates. A lower prevalence was observed for C. jejuni with seven identified isolates and one isolate was identified as C. helveticus. Multi-locus sequence typing (MLST) was carried out on the seven C. jejuni isolates and all sequence types that were found are also commonly found in humans. The dogs were divided into three age groups; 1) under 12 months, 2) 12 to 23 months and 3) 24 months and older. The highest prevalence was found in the two younger age groups. Dogs shedding C. jejuni were between 3–12 months of age while dogs shedding C. upsaliensis were found in all ages.

Conclusions

The present investigation finds that Campylobacter spp. known to cause campylobacteriosis in humans are present in Swedish dogs. The results suggest an age predisposition where dogs under 2 years of age are more likely to shed Campylobacter spp. than older dogs. The most commonly isolated species was C. upsaliensis followed by C. jejuni, which was only detected in dogs up to 12 months of age. All C. jejuni isolates identified in the present study were of the same MLST types that have previously been described both in humans and in animals. The awareness of the Campylobacter risk of healthy young dogs may be an important way to reduce the transmission from dogs to infants, young children and immunocompromised adults.     

Pathology of fatal highly pathogenic H5N1 avian influenza virus infection in large-billed crows (Corvus macrorhynchos) during the 2004 outbreak in Japan

Highly pathogenic H5N1 avian influenza viruses were isolated in 9 large-billed crows that died in Kyoto and Osaka prefectures in Japan from March to April in 2004. We studied 3 of the 9 crows using standard histologic methods, immunohistochemistry, and virus isolation. The most prominent lesions were gross patchy areas of reddish discoloration in the pancreas. The consistent histologic lesions included severe multifocal necrotizing pancreatitis, focal degeneration and necrosis of neuron and glial cells in the central nervous system, and focal degeneration of cardiac myocytes. All of these tissues contained immunohistochemically positive influenza viral antigens. The virus was isolated from the brain, lung, heart, liver, spleen, and kidney of the crows examined. Thus we concluded that highly pathogenic avian influenza virus was associated with clinical disease, severe pathologic changes, and death in the 3 crows.     

Antimicrobial susceptibilities, serogroups, and molecular characterization of avian pathogenic Escherichia coli isolates in Japan

In total, 83 avian pathogenic Escherichia coli (APEC) isolates from avian colibacillosis during a period from 2001 to 2006 in Japan were investigated for serogroups, typical virulence factors, antimicrobial susceptibility, and genetic relatedness. The most common serogroup was O78 (30.1%); 80.7% of isolates harbored the iss gene and 55.4% of isolates harbored the tsh gene. Antimicrobial resistance of the isolates was found for ampicillin (77.1%), oxytetracycline (75.9%), kanamycin (36.1%), fradiomycin (33.7%), trimethoprim (25.3%), enrofloxacin (21.7%), and florfenicol (6.0%). Although multiple antimicrobial-resistant phenotypes (three or more antimicrobials) accounted for 54.2% of isolates, no isolate exhibited resistance to all agents tested. The fluoroquinolone-resistant isolates had point mutations in GyrA (Ser83 --> Leu, Asp87 --> Asn) and ParC (Ser80 --> Ile, Glu84 --> Gly). Of 18 enrofloxacin-resistant E. coli isolates, nine isolates belonged to serotype O78. In PFGE analysis, eight of the nine enrofloxacin-resistant O78 isolates were classified into an identical cluster. This suggests that a specific genotype of fluoroquinolone-resistant O78 APEC may be widely distributed in Japan.     

Retinal Photoreceptor Fine Structure in the American Crow (Corvus brachyrhynchos)

The morphology of the light-adapted retinal photoreceptors of the American crow (Corvus brachyrhynchos) has been investigated by light and electron microscopy. They consist of rods, single cones and double (unequal) cones present in a ratio of about 4:3:3 respectively. The rods are stout cells with a long inner segment and an outer segment that reaches to the RPE cell body. In the light-adapted state, the pigment-laden apical processes of the RPE cells surround cell photoreceptor types for most of their length. The rod inner segment displays an ellipsoid of mitochondria, a large hyperboloid of glycogen, much RER, numerous polysomes, Golgi zones and autophagic vacuoles. Single cones show a slightly tapered outer segment, a large and usually heterogeneous oil droplet and an ellipsoid of mitochondria at the apex of the inner segment. Double cones consist of a longer, stouter chief member which displays a more homogeneous oil droplet and a prominent paraboloid of glycogen and a slightly shorter and thinner accessory member with no oil droplet or paraboloid. Both members of the double cone as well as the single cones show a prominent ellipsoid and plentiful polysomes, RER and Golgi zones in the inner segment. Along the length of the contiguous membranes of the two members of the double cone are presumed interreceptor junctions. All cone photoreceptors are relatively small in diameter and hence are tightly packed. Judging by their morphology in the light-adapted state neither rods nor cones are felt to undergo photomechanical movements in this species. Rods and cones (both types) display both invaginated (ribbon) synapses as well as numerous flat (conventional) synaptic sites.     

Fine Structure of the Pecten Oculi in the American Crow (Corvus brachyrhynchos)

The pecten oculi of the American crow (Corvus brachyrhynchos) has been examined by both light and electron microscopy. In this species the pecten is very large and of the pleated type. It consists of 22 - 25 accordion folds that are joined apically by a bridge of tissue which holds the pecten in a fan-like shape widest at its base. Within each fold are numerous capillaries, larger supply and drainage vessels and many melanocytes. The capillaries are extremely specialized for transport functions and display extensive microfolds on both their luminal and abluminal borders. Except for the nuclear region which contains most of the organelles, the endothelial cell bodies are extremely thin. These capillaries are surrounded by thick fibrillar basal laminae which are conjectured to be structurally important. Pericytes are a common feature of these capillaries. The numerous pleomorphic melanocytes interspersed between the capillaries are also felt to be important in structural support of the pecten. The pecten is considered to be comparable to the falciform process of some teleosts, the conus papillaris of reptiles, the supraretinal vessels of amphibians and teleosts and the intraretinal vessels of mammals which are all felt to be alternative methods of bringing nutrients to the inner retina.