Newcastle disease virus (NDV) vectors expressing avian influenza virus (AIV) hemagglutinin of subtype H5 protect specific pathogen–free chickens from Newcastle disease and avian influenza. However, maternal AIV antibodies (AIV-MDA+) are known to interfere with active immunization by influencing vaccine virus replication and gene expression, resulting in inefficient protection. To overcome this disadvantage, we inserted a transgene encoding a truncated soluble hemagglutinin (HA) in addition to the gene encoding membrane-bound HA from highly pathogenic avian influenza virus (HPAIV) H5N1 into lentogenic NDV Clone 30 genome (rNDVsolH5_H5) to overexpress H5 antigen. Vaccination of 3-wk-old AIV-MDA+ chickens with rNDVsolH5_H5 and subsequent challenge infection with HPAIV H5N1 3 wk later resulted in 100% protection. Vaccination of younger chickens with higher AIV-MDA levels 1 and 2 wk after hatch resulted in protection rates of 40% and 85%, respectively. However, all vaccinated chickens showed strongly reduced shedding of challenge virus compared with age-matched, nonvaccinated control chickens. All control chickens succumbed to the HPAIV infection with a grading in disease progression between the three groups, indicating the influence of AIV-MDAs even at a low level. Furthermore, the shedding and serologic data gathered after immunization indicate sufficient replication of the vaccine virus, which leads to the assumption that lower protection rates in younger AIV-MDA+ chickens are caused by an H5 antigen–specific block and not by the interference of the AIV-MDA and the vaccine virus itself. In summary, solid protective efficacy and reduced virus transmission were achieved in 3-wk-old AIV-MDA+ chickens, which is relevant especially in regions endemically infected with HPAIV H5N1.

Vaccine-related fowl cholera must be considered when flock mortality increases after use of a live Pasteurella multocida vaccine product. All registered live vaccines serotype as Heddleston 3,4; however, in some regions this is also the most common serotype of outbreak isolates in broiler breeders and turkeys. Therefore, serotyping may not be useful for diagnosing vaccine-related fowl cholera. This project sought to apply a vaccine-specific test to differentiate vaccine-related disease from naturally occurring outbreaks. Results indicate that vaccine strains were commonly isolated from broiler breeders exhibiting signs of fowl cholera postvaccination, but some of these isolates exhibited only serotype 4 antigenicity. The isolates' lipopolysaccharides, the target antigen for serotyping, contained compositional changes that may explain the varying serotype results and virulence of the commercial preparations. These results suggest that vaccine-related disease may be common in broiler breeders, and live commercial vaccine preparations need to be assessed for serotype and titer prior to use in order to reduce vaccine-related fowl cholera.

Many H5 and H7 subtype avian influenza vaccines are poorly immunogenic in terms of inducing hemagglutination-inhibition (HI) antibody titers. Residue 227 (H3 numbering) in the receptor binding site in the hemagglutinin (HA) is critical for the detectability of HI antibodies induced by H5 influenza vaccines. However, whether the effect of residue 227 on immunogenicity can be generalized in different subtypes is unclear. In this study, the impact of HA residue 227 on immunogenicity of H5N1, H5N6, and H7N9 avian influenza vaccines was evaluated in chickens. Polymorphism analysis revealed that S227 is overwhelmingly dominant in HA of the H5N1 and H7N9 subtypes, whereas this amino acid is present in a small proportion of H5N6 viruses. The H5N1, H5N6, and H7N9 vaccines harboring S227 in HA induced relatively low HI titers at week 2 postimmunization (pi), and antibody titers increased at week 3 pi. S227N substitution in these vaccines consistently enhanced HI titers significantly. Another H5N6 vaccine harboring Q227 in HA elicited a robust HI antibody response, and Q227S substitution led to a significant drop of HI titers. Cross-HI testing against the wild-type and mutant viruses revealed that the amino acid at position 227 was associated with the detectability of HI titers induced by H5 and H7 avian influenza vaccines. The results indicate an important role of residue 227 in HA in immunogenicity of H5 and H7 subtype avian influenza vaccines in chickens. Our findings also provided useful information for vaccine seed virus selection and genetic engineering for immunogenicity enhancement of avian influenza vaccines.

Major histocompatibility complex (MHC) congenic chicken lines have been used as a model to study infectious bronchitis virus (IBV) immune responses in chickens. Zinc (Zn) and manganese (Mn) are trace minerals that act as enzyme cofactors in cellular reactions. In addition, Zn is an important modulator of immune responses, especially in the respiratory tract. Zinc and Zn + Mn amino acid complex supplements were tested to alleviate the effects of an IBV challenge using relatively resistant and susceptible MHC congenic chicken lines. Prior to the challenge with IBV, the amino acid–bound supplements induced better weight gain in the IBV-resistant chicken line (331/B2) compared to the birds fed with the sulfate-delivered supplements. No body weight differences were detected between IBV-challenged and unchallenged 331/B2 birds supplemented with Zn in amino acid complex. A reduction of respiratory signs was observed in 335/B19 birds fed with the diet supplemented with Zn in amino acid complexes at 4 dpi. Compared to the sulfate-bound trace minerals, 331/B2 chickens fed with the amino acid–bound supplements presented milder clinical sign trends at 6 dpi and less severe airsacculitis at 14 dpi. The total antibody response in serum in 331/B2 birds fed with the amino acid–bound Zn ration was the highest among all groups tested. Both amino acid–delivered trace mineral supplements induced a slightly higher antibody response than the sulfate-bound ration in both chicken lines. This experiment provides insights into the effect of Zn and Mn on the immunity of chickens with known different susceptibilities to IBV.

Zebra finches (Taeniopygia guttata) are laboratory animal species commonly used for modeling neurobiology and learning. Historically, using bacterial culture, biochemical analysis, and 16S ribosomal RNA gene sequencing, bacterial isolates from feces of finches housed at Massachusetts Institute of Technology had been presumptively diagnosed as Campylobacter jejuni, which is commonly isolated from both domestic and wild birds. Although the zebra finches were not clinically affected, C. jejuni is a known zoonotic pathogen that causes gastroenteritis in humans worldwide. Human transmission is predominantly foodborne and associated with the consumption of contaminated poultry; however, humans can also become infected from contact with C. jejuni-infected reservoir hosts. Because C. jejuni-infected finches pose a risk to research personnel, a study was undertaken to investigate the prevalence and taxonomic identification of Campylobacter spp. present in the finch colony. Campylobacter spp. were isolated from a total of 26 finch fecal samples collected in 2003, 2010, and 2017. 16S ribosomal RNA sequencing of all isolates determined that they shared 99% identity with either C. jejuni or Campylobacter lari. Sixteen of the isolates were subjected to further biochemical characterization and atpA and rpoB gene sequence analysis. Based on these analyses, three clusters of Campylobacter species were identified. The draft whole-genome sequences were determined for one representative isolate from each cluster. A pan-genomic phylogenetic tree, average nucleotide identity, digital DNA-DNA hybridization, and orthologous gene analyses indicated that each isolate was its own novel species, distinct from C. jejuni and other avian Campylobacter species. We have named these novel species Campylobacter taeniopygiae, Campylobacter aviculae, and Campylobacter estrildidarum, and in each novel species, we identified virulence genes suggesting their pathogenic and zoonotic potential.

Salmonella enterica serotypes Enteritidis (SE) and Heidelberg (SH) are consistently linked to poultry-related foodborne outbreaks and can be isolated from broiler parts in processing facilities. In order to control this pathogen's establishment in the broiler, entryways at the farm that lead to colonization must be considered. The objective of these trials was to determine if the inoculation route of either SE or SH altered its recovery in a market-age broiler's digestive tract if chicks were dosed on day of hatch. Chicks were given a 10⁴ colony-forming units inoculation of SE or SH on day 0 via one of five inoculation routes (oral, intratracheal, subcutaneous, ocular, or cloacal) and then placed in pens (60–100 chicks/treatment). Broilers were reared for 32–36 days, then euthanatized, and samples of trachea, crop, liver and spleen (pooled), cecum, and a cloacal swab were collected. Samples were enriched and then analyzed on yes/no criteria based on Salmonella growth. Data were analyzed in JMP Pro 14.1 using the GLM procedure with the Student t-test to separate serotype means and a Tukey honestly significant difference test to separate inoculation means (P ≤ 0.05). All samples collected and all inoculation routes resulted in recovery of either serotype. The intratracheal inoculation, mimicking inhaled fomites, resulted in significantly higher recovery of Salmonella serotypes than did the other inoculation routes (P < 0.0001), indicating the importance of controlling respiratory contamination. When comparing serotypes, there was a significantly greater recovery of SH compared to SE based on samples collected (P = 0.001). SH also had significantly greater recovery from the cecum (P < 0.001) and the cloacal swab (P = 0.02). These trials indicate the need for further investigation of the intratracheal route, as well as reinforcing that the potential of systemic infection through grow out with either serotype is highly probable preharvest.

Since August 2014, the University of Minnesota Veterinary Diagnostic Laboratory has received cases of turkey enteritis that are clinically different from previously described cases of poult enteritis syndrome and light turkey syndrome. The birds develop dark green and extremely foul-smelling diarrhea starting at 8–10 wk of age, which may last up to 15–16 wk of age. The affected turkey flocks show poor uniformity, and feed conversion and market weights are reduced. Multiple-age farms are affected more often than the single-age farms. Morbidity varies from flock to flock and in some cases reaches 100%. At necropsy, undigested feed with increased mucus is observed in the intestines along with prominent mucosal congestion and/or hemorrhage. Microscopically, lymphocytic infiltrates expand the villi in duodenum and jejunum to form lymphoid follicles, which are often accompanied by heterophils. Next generation sequencing (Illumina Miseq) on a pool of feces from affected birds identified genetic sequences of viruses belonging to Astroviridae, Reoviridae, Picornaviridae, Picobirnaviridae, and Adenoviridae. On testing pools of fecal samples from apparently healthy (16 pools) and affected birds (30 pools), there was a higher viral load in the feces of affected birds. Picobirnavirus was detected only in the affected birds; 20 of 30 pools (66.7%) were positive. These results indicate that a high viral load of turkey picobirnavirus alone, or in association with novel picornaviruses, may be a cause of this new type of turkey enteritis.

An adult blue-fronted Amazon parrot (Amazona aestiva) was presented for a 6-wk history of ataxia and weight loss. Complete blood count, plasma chemistry panel, bile acids, and radiographic imaging were considered normal or unremarkable. The patient was hospitalized and supported with subcutaneous fluids, vitamin B complex, meloxicam, enrofloxacin, gavage feeding, and fenbendazole. While hospitalized, the ataxia significantly improved, and the bird began eating on its own and gaining weight. The bird was discharged from the hospital and prescribed enrofloxacin, meloxicam, and fenbendazole to be administered by the owner with recommendations for routine follow-up care. Medications were discontinued before emergent representation; at the time of reevaluation, the patient's condition had deteriorated severely. Given the poor prognosis, the owners elected for euthanasia. No gross abnormalities were noted on postmortem examination. Liver tissue zinc levels measured 125 ppm; normal limit is less than or equal to 25 ppm. Histopathologic changes to the brain were consistent with severe zinc toxicosis demonstrated by vasculopathy of the cerebral arteries and arterioles with multifocal areas of hemorrhage and astrocyte swelling. These findings have been reported in humans and other mammals but not birds. Although the source of this bird's heavy metal exposure is unknown, the high tissue zinc concentrations imply chronic exposure. This case presentation and unusual pathologic findings will be beneficial to the further understanding of avian zinc toxicosis.

In March 2019, the California Animal Health and Food Safety Laboratory (CAHFS), Turlock branch, received two submissions of broiler chickens from commercial flocks reporting increased mortality. Submissions consisted of either white or brown broilers. Submitted chickens appeared depressed with ruffled feathers. At necropsy, moderate to severely enlarged and pale kidneys were observed, with gross lesions indicative of dehydration. Microscopically, renal tubules were degenerated and distended with necrotic debris and tubular casts. The kidney parenchyma contained mononuclear inflammatory cell infiltrates and interstitial edema. Infectious bronchitis virus (IBV) was isolated and identified by reverse transcription quantitative PCR from kidney tissue pools and tracheal swab pools from both cases. Partial sequencing of the  S1 (482_avdi-64-04-03_s01.doc) hypervariable region was most similar to a local California variant, CA1737. The outbreak lasted roughly 1 wk in both flocks, with 2% total mortality in the brown broilers and 20% total mortality in the white broilers. Final proof of the IBV strains causing nephropathy will require fulfillment of Koch postulates. IBV associated with nephropathy has been sporadically reported in California chicken flocks and represents a significant pathogen due to its potential for inducing high flock mortality. The incidence of IBV associated with a nephropathy diagnosis in chicken necropsy submissions to the CAHFS system-wide from 1998 to 2019 is also reviewed.

Necrotic enteritis (NE) is a devastating enteric disease caused by Clostridium perfringens type A/G, which affects global poultry industry by compromising the performance, health, and welfare of chickens. The causative main virulent factor responsible for NE pathogenesis has been shifted from a phospholipase C portion of an α-toxin, to an NE B-like (NetB) toxin, a plasmid-encoded pore-forming heptameric protein, in NE development. Therefore, the ability to detect NetB toxin will enable early diagnosis of field NE. Because the NetB protein can only be detected by western blot analysis with polyclonal anti-NetB antiserum, we developed a NetB-specific monoclonal antibody (mAb)–based capture enzyme-linked immunosorbent assay (ELISA). Twenty mAbs reacting with Escherichia coli–expressed NetB protein were selected, isotyped, and conjugated with horseradish peroxidase for antibody pair tests. Multiple mAb pairs were found to detect E. coli NetB protein and native NetB protein secreted by netB-positive C. perfringens isolates. The developed capture (sandwich) ELISA could be useful to identify in vitro production of native NetB protein secreted from netB-positive field C. perfringens isolates and to conduct a large field test of commercial chickens undergoing NE infection. Here, we first report that native NetB toxin can be detected in C. perfringens NetB-specific mAb-based capture ELISA.

The avian pathogen Ornithobacterium rhinotracheale (ORT) has been implied in the etiology of poultry respiratory disease in recent years. To evaluate whether Whatman^® Flinders Technology Associates (FTA^®) cards can be used for hazard-free transport and storage of ORT samples for posterior DNA amplification, a controlled assay was performed. Three 10-fold dilutions of an ORT culture suspension were spotted on FTA cards and stored at room temperature (RT) for 6 mo. Sterile swabs were immersed in the same three 10-fold culture dilutions and stored at RT and 4 and –20 C without storage medium for the same time. DNA was extracted from both the FTA cards and swabs 1 day, 1 and 6 wk, and 6 mo following sample preparation and stored at –20 C. At the end of the experiment, real-time PCR amplification of the 16S ribosomal RNA gene was performed from DNA extracted throughout a 6-mo period from all ORT samples stored on both FTA cards and swabs. The obtained threshold cycle values for each ORT DNA extraction date were within the same range for all samples in a dilution-dependent fashion, regardless of storage temperature or used material. Pure ORT colonies could be reisolated 1 day after sample preparation from the swab dilutions stored at all temperatures but not from the FTA cards. We conclude that the efficiency of ORT DNA amplification from samples stored on FTA cards or in swabs is similar. However, FTA cards have the advantage of preventing microorganism growth, thus allowing safe transport and storage, for at least 6 mo, for bacterial dilutions down to at least 10⁴–10⁵ colony-forming units/ml.

Erysipelas is a bacterial disease caused by Erysipelothrix rhusiopathiae that affects multiple mammalian and avian species. In poultry, the disease is of sporadic prevalence and more often observed in older birds, leading to decreased egg production and mortality. Among avian species, turkey breeders seem to be the most affected, but outbreaks have been reported in ducks, layer chickens, quails, geese, and various captive and free-range birds. Sixty-seven cases of erysipelas have been diagnosed in animals submitted for necropsy evaluation at the California Animal Health and Food Safety Laboratory System from January 2000 to December 2019. Of these, 38 cases (56.72%) were in avian species, and a retrospective analysis of these avian cases was performed. The majority of the avian cases were in turkeys (17/38, 44.74%). Most of the turkey breeder cases reported performing artificial insemination prior to the increase in mortality. In other birds, mortality was often observed without observing previous clinical signs. The majority of cases presented with coinfections with other pathogens (23/38, 60.53%), which might have affected the clinical outcome. Despite the occasional occurrence in avian species, erysipelas is an important pathogen in poultry and should be considered as a differential diagnosis in other avian species when acute septicemia is suspected as the cause of mortality.

Mycoplasma gallisepticum (MG) is a major pathogen of the poultry industry throughout the world. MG causes chronic respiratory disease in chickens and infectious sinusitis in turkeys. Despite constant improvements in the biosecurity of the poultry industry in Iran, MG infection still occurs and causes significant economic issues. To evaluate genetic variability, 10 Iranian MG isolates along with 17 available sequences were characterized by gene-targeted sequencing (GTS) analysis of complete mgc2/pvpA genes. According to the findings, 21 different sequence types within the sample set of 27 strains were typed by this method. The discriminatory power of this typing assay was established to be 0.97. Although no insertions and deletions of nucleotides were observed in the mgc2 gene among the Iranian strains, different lengths of pvpA genes with 1086, 1095, and 1101 nucleotides were detected within direct repeats (DRs) 1 and 2. Generally, eight tetrapeptides Pro-Arg-Pro-Met/Gln/Asn were found in the DRs of PvpA. Analysis of the carboxyl ends of PvpA proteins exhibited various repeats of prolines. In the phylogenetic tree of partial and complete mgc2/pvpA genes, all Iranian MG isolates were clustered into two distinct groups. Because this typing assay could provide a higher discriminatory power than the previously reported GTS scheme of partial mgc2/ pvpA genes, these results can be considered a blueprint for future national control and diagnostic strategies. Furthermore, consistent surveillance with larger datasets will be needed to clarify the epidemiologic characteristics of MG outbreaks in different poultry hosts.

A game bird producer in the North Central region of the United States submitted unhatched ring-neck pheasant (Phasianus colchicus) eggs for diagnostic evaluation. The submitting complaint was a drastic drop in hatchability. This operation has its own breeder birds that are housed in outside pens. This hatch occurred in the latter third of the production cycle. Typical hatchability for this operation is around 75% (± 3%). The hatchability of this hatch was between 14%–15%. Approximately 30,000 eggs were set with an expected hatchability of about 23,000 birds. The number of birds from this hatch was less than 4500, with a net loss approaching 20,000 chicks. All unhatched eggs submitted were in late stage development. The chick embryos had pipped through the shell but died before hatching. Approximately 5000 eggs originating from an outside breeder source were also set at the same time in the same machines and experienced a normal hatch. The exterior surfaces of the eggshells of the unhatched eggs experiencing low hatchability were swabbed and submitted for bacteriologic evaluation. Additionally, embryos from some of the unhatched eggs were removed aseptically from their eggshells, and their internal organs were harvested and submitted for bacteriologic evaluation. The bacteriology results identified no pathogenic bacteria from the eggshells. However, the embryo samples revealed large quantities of Enterococcus faecalis. In discussions with the producer, the only factor identified was an unusually warm period followed by an atypically cold and wet period during the time of egg collection for those eggs experiencing low hatchability.

Field visits at two different farms suggest a correlation between commercial turkey (Meleagridis gallopavo) flocks having increased mortality from blackhead disease (histomoniasis) if they suffer from poor poult quality at placement and coccidiosis (Eimeria spp.) before age 6 wk. In both cases, the flocks were all-in/all-out with curtain-sided houses and received a coccidiosis vaccine on day of hatch. At Farm I 2018, poults from different hatcheries were placed in two houses on the same farm (Houses 1 and 2). House 2 had poults considered poor quality and suffered from mortality associated with coccidiosis at 2 and 4 wk of age. At 8 wk, blackhead disease was diagnosed in both houses by postmortem examination. House 2 had mortality of >2000 poults, and the subpopulation of necropsied poults had gross lesions characteristic of histomoniasis. Gross lesions associated with blackhead disease were only found in eight poults in House 1, which was populated with good-quality poults and did not have a second spike in mortality due to coccidiosis. The Farm II 2020 poults were delivered from the same hatchery onto a three-house farm (Houses A, B, and C). House C had poults that were considered poor quality and had mortality associated with coccidiosis at 3 wk of age. At 8–9 wk, House C had mortality approaching 1000 birds, with all poults examined postmortem having clinical signs of blackhead disease. Houses A and B were populated with good-quality poults and had no diagnosed mortality from coccidiosis or blackhead disease. The similarity of these two cases suggest that poult quality at placement coupled with coccidiosis before 6 wk of age can influence the severity of blackhead disease in commercial turkey flocks.

Transmissible viral proventriculitis (TVP) is a disease of chickens, mostly in broilers of 2–8 wk of age. Chicken proventricular necrosis virus (CPNV), a birnavirus, is the etiologic agent. Characteristic gross lesions are enlargement, atony, and pallor of the proventriculus. Cases diagnosed in California between 2000 and 2018 (n = 477), originating from 93 different farms representing all major companies in the region, were analyzed. Frequency of cases varied widely between years, with no recognizable seasonality. The flocks were between 6 and 61 days of age; the average age was 34.0 days, and the median age was 35 days. In 166 cases, between 6.3% and 100% of the submitted birds had gross lesions in the proventriculus. The most common findings were enlarged or dilated proventriculi, thickened walls, and pale or mottled serosal appearance. Histopathologically, inflammation of the glands was the most frequent finding. Other lesions included necrosis, hyperplasia, or both conditions of the glandular epithelium; dilated glands; and occasionally fibrin deposition, fibrosis, and hemorrhages. Twenty-three proventriculi from six cases were tested by immunohistochemistry for the presence of CPNV antigen; 21 stained positive. In 209 cases, birds also had lesions in the bursa fabricii attributed to infectious bursal disease, but with no significant difference in the mean percentage of birds with gross lesions in the proventriculus between cases with or without lesions in the bursa fabricii. The results show that TVP is a common disease of broiler flocks in California and confirms that CPNV is the likely causative agent.

This study was performed to evaluate the diversity and prevalence of yeasts associated with esophageal mycosis in domestic ducks and geese. Fungi were isolated from esophageal lesions of dead animals sent for microbiologic laboratory diagnosis. Species identification using a culture-dependent method was carried out by sequencing of the internal transcribed spacer (ITS)1-5.8S rRNA-ITS2 region. The most frequently isolated yeast was Candida albicans (43.1%) followed by Saccharomyces cerevisiae (17.6%), Candida kefyr (11.7%), Kazachstania bovina (11.7%), Candida lambica (3.9%), and single isolates (1.9%) representing Candida inconspicua, Candida rugosa, Candida pelliculosa, Candida krusei, Magnusiomyces capitatus, and Trichosporon asahii. Our results indicate that a number of potentially pathogenic yeast species can be isolated from esophageal mycosis of waterfowls, but additional studies are needed to make conclusions regarding their possible etiologic role in disease.

The prevalence of Gallibacterium anatis in poultry production has increased over the last two decades. However, only a few studies have explored the pathogenicity of this bacterium in commercial layer chickens. This trial studied the aspects of the pathogenicity of a Gallibacterium anatis biovar haemolytica local Egyptian isolate (previously registered as strain B14 with GenBank accession no. KJ026147). We used 500 base pairs of a 16S ribosomal RNA gene and the 16S-23S ribosomal RNA intergenic spacer, partial sequence in an experimental infection trial in commercial White Shaver layer chickens aged 19 wk. The hens were divided into three groups of 40 birds each. The hens in Groups 1 and 2 were experimentally infected through the intranasal (IN) and intravenous (IV) routes, respectively, with a dose of 0.2 ml/bird containing 1.2 × 10⁹ colony-forming units/ml. In contrast, Group 3 was kept as a noninfected control group. Both IN and IV infections resulted in a delayed egg laying for 1 wk and a significant (P ≤ 0.05) drop in egg production by 7.81% and 10.28% compared with the control group over 7 wk. Severe lesions in the form of hemorrhagic pneumonia, catarrhal tracheitis, ovarian follicle and oviductal regression, and septicemia were evident on necropsy, demonstrating the pathogenicity of G. anatis as a primary pathogen.

Avian chlamydiosis is an infection caused by obligate intracellular, gram-negative bacteria belonging to the Chlamydiaceae family. Birds can be hosts of several Chlamydia species, including Chlamydia avium, which has only been detected in pigeons and psittacine birds. In this study, depression, respiratory distress, and mortality were noted among psittacines belonging to a large aviary with 35 different avian species. On the basis of immunohistochemistry and PCR testing, chlamydiosis was diagnosed in affected birds. Gross and histopathologic lesions were mainly observed in the spleen and gastrointestinal tract. Chlamydia avium was detected in four psittacines by PCR, including two dead birds and two individuals exhibiting respiratory distress. Increased aspartate aminotransferase and lactate dehydrogenase values and anemia were consistently identified in affected birds. Administration of doxycycline, combined with hepatoprotectors and vitamins, was effective in stopping mortality and bacterial shedding.

Tetratrichomonas is a genus of parasites that usually inhabits the lower digestive tract, especially the cecum, of various bird species. The infection might lead to birds' death, but in many cases it could be asymptomatic or with mild clinical signs which might be not observed. Subclinical infections can be undiagnosed, leading to production losses. To investigate the prevalence of Tetratrichomonas spp. in geese, 23 cloacal swabs were taken from each of 43 flocks of reproductive geese from five major geese production provinces in Poland after first, second, or third laying season. The obtained swabs were placed in culture medium for propagation of the parasite. All cultures were screened microscopically before PCR was applied to detect the parasites' DNA. After cultivation, the presence of genetic material of Tetratrichomonas was found in 430 out of 989 samples, which correspond to 38 (88.4%) of 43 flocks. The study shows how the number of laying seasons and the size of the flock in which the birds were kept affects the distribution of protozoa of the genus Tetratrichomonas in geese reproductive flocks in Poland.

The prevalence of Heterakis infection in reproductive geese from 56 flocks was investigated between February 2015 and November 2019 by using both anatomopathologic and coproscopic examinations. The nematodes were microscopically examined, and 37.5% (21/56) of the flocks were infected with Heterakis nematodes, mostly with Heterakis dispar (35.7%). In one particular flock, a Heterakis gallinarum infection was noted. In two flocks, Heterakis infection was identified only during coproscopic examination. The infection susceptibility of the geese depended on the bird's age—in the first reproductive season, 50% of the flocks were positive; in the second season, 42% were positive; and in the third season, 30% were positive. The number of nematodes in each necropsied goose varied from 8 to 216 individuals. During the 5 years of epidemiologic analysis, the infection occurrence decreased from 66.7% to 11%.

In the spring of 2019, adult (75 wk old) brown laying hens from a commercial, pen-free, egg-laying facility (11,000 birds per house) located in northwest Arkansas were obtained for the purposes of sourcing robust and evenly dispersed cestode infections for anticipated anthelmintic evaluations. To that end, four birds from each of six discrete sites (northwest, northcentral, northeast, southwest, southcentral, and southeast) in one production barn were obtained on two occasions, 8 days apart, and necropsied for helminth counts. A definite, repeated, location-to-location variation in infection incidence and magnitude was seen for each of the two cestode parasite species present and for one of the two nematode parasite species present. Burdens of Ascaridia galli were generally similar regardless of bird location, with site-specific mean totals per bird over both sampling days between 31 and 80. For the remaining helminths, infections were greatest for birds from the southern half of the building as opposed to the northern and from the western end of the barn as opposed to the central or eastern portions. Location-specific mean worm burdens over both sample dates ranged from 340 to 1133 (Heterakis gallinarum), 14 to 277 (Raillietina cesticillus), and 1 to 35 (Choanotaenia infundibulum). The greatest individual bird infections were 299 (A. galli), 3575 (H. gallinarum), 1015 (R. cesticillus), and 102 (C. infundibulum). The above counts are for all developmental stages combined (A. galli and H. gallinarum) and for scolexes only (R. cesticillus and C. infundibulum), as determined via standard collection and quantification procedures using both intestinal contents and overnight soaks. Immediately before the mapping study outlined above, birds were obtained from the east end of the source barn and used for the nematocidal evaluation of fenbendazole in the water (5 mg/kg body weight [BW] for 1 day), levamisole in the water (8 mg/kg BW for each of 2 days), herbal mixture in the feed (1 gm/4.5 kg BW each day for 5 days), diatomaceous earth (2% of total feed for 10 days), and a nutraceutical mixture feed supplement (2% of feed for 7 days). Based on arithmetic means for adult forms, control trial efficacies for A. galli and H. gallinarum were 0% and 12% for the nutraceutical feed additive, 0% and 22% for the diatomaceous earth feed additive, 0% and 26% for dietary herbals, 87% and 63% for levamisole, and 82% and 84% for fenbendazole, respectively. Only adult burdens of A. galli and H. gallinarum for fenbendazole- and levamisole-treated birds were significantly different from control bird levels (P ≤ 0.05).

Over a 4-mo period, a Michigan zoo had 32 budgerigars, Melopsittacus undulatus, from their flock die. Whole animals or formalin-fixed tissues were submitted to Michigan State University Veterinary Diagnostic Laboratory for diagnosis. Avian gastric yeast infection, Macrorhabdus ornithogaster, was diagnosed in seven birds. There was atrophy of breast musculature and no subcutaneous or coelomic fat stores in six necropsied birds. Only two birds had proventricular dilatation grossly. Histologic examination of the proventriculus of all seven birds revealed abundant 3 × 50-µm septate, parallel-walled, nonbranching yeast organisms morphologically consistent with Macrorhabdus ornithogaster. Mycobacteriosis was diagnosed in three budgerigars, two of which were necropsied. Both necropsied birds had hepatomegaly and one also had splenomegaly. No acid-fast bacilli were found in the livers of either bird but were found in splenic macrophages of the bird with splenomegaly and in the intestine of the other bird. Mycobacterium species were cultured from the enlarged spleen and identified by DNA sequence as Mycobacterium genavense. Pulmonary granulomas with acid-fast bacilli were found in the bird submitted as fixed tissues. None of the budgerigars had a dual infection. The remainder of the budgerigars died from hepatitis, nephrosis, oviductal prolapse, exclusion from food and water by flock mates, or tumors.

Dust collected from the poultry house has been increasingly used as a population-level sample to monitor the presence of pathogens or to evaluate the administration of live vaccines. However, there are no guidelines for the storage of this sample type. This study investigated the stability of infectious laryngotracheitis virus (ILTV), a DNA virus, and infectious bronchitis virus (IBV), an RNA virus, in poultry dust kept under temperature and moisture conditions that mimic on-farm and laboratory storage. Dust samples were collected from chicks spray vaccinated with a live IBV vaccine and inoculated with a field ILTV strain via eye drop. Samples were stored under different moisture conditions (dry = 2% moisture, moist = 22%–71% moisture) and temperatures (–20, 4, 25, and 37 C) for different durations (0, 7, and 14 days, and 1, 2, 3, and 4 mo) in a factorial arrangement, followed by quantitative PCR for detection of virus genome copies (GC). The length of storage, moisture level, and storage temperature affected the viral genome load for ILTV and IBV but did not affect the number of positive samples for each virus. All treatment combinations were ILTV positive for at least 4 mo. In dry dust samples, all storage temperatures or durations had quantifiable ILTV or IBV GC. Moisture addition had a detrimental effect on viral genome load, causing an overall reduction of 0.3 log ₁₀ for ILTV GC (7.29 and 6.97 log ₁₀, P = 0.0001), and 1.3 log ₁₀ for IBV GC (5.95 and 4.66 log ₁₀, P = 0.0001), which are unlikely to have biologic significance. In conclusion, dry dust can be stored at any temperature up to 37 C for at least 4 mo without loss in qPCR detection of ILTV or IBV GC. Collection or storage of moist dust should be avoided, or air drying prior to storage is recommended if only moist dust is available.

Histologic and bacteriologic features for groups of average 31-day-old broilers displaying three gross categories of femoral head alterations were documented. Categories included simple femoral head separation (FHS), femoral head transitional changes (FHT), and femoral head necrosis (FHN). Groups with grossly normal (NORM) femoral heads and cull birds with FHN and having gross signs of sepsis (Cull-FHN) were also included in the study. There was a 10% occurrence of positive bacterial cultures for all birds tested. Most positive cultures (33%) were found in the Cull-FHN group, while only a 12% occurrence was seen in the FHS group, and no positives were present in the FHT or FHN groups. A 14% total occurrence of femoral bacterial chondronecrosis with osteomyelitis or simple osteomyelitis (BCO-O) was observed. A progressive increase in the prevalence of BCO-O was apparent between groups going from NORM (0%), FHS (4%), FHT (14%), FHN (13%), and reaching a maximum of 67% in the Cull-FHN group. Minimal to mild femoral head cartilage necrosis was present in 40% of NORM broilers and 100% of the FHS, FHT, and FHN groups, but at moderate severity in 20% of the Cull-FHN group. Thus, the majority of FHN cases were associated with aseptic cartilage necrosis rather than BCO-O. These findings suggest that aseptic cartilage necrosis may be as important as septic necrosis as a cause of gross femoral head disease. A 26% overall occurrence was seen for hip synovitis–arthritis, but group differences were not statistically significant. Synovitis was not seen in the NORM group and was present in some (12%) of the FHS group but was observed at a high rate in both the FHN (43%) and the Cull-FHN (50%) groups. Morphometric measurements demonstrated that the area size of femoral fibrous cortical defects or “cutback zones” were generally larger for all gross categories relative to NORM, with a significant difference between NORM and FHS groups. This study underscores the multifactorial etiology of FHN and the importance of conducting both histologic and bacteriologic evaluations in which gross evidence of FHN or BCO-O occurs.