Infection with Cochlosoma anatis, a flagellated protozoan parasite, significantly impacts U.S. commercial turkey production by causing poult enteritis (poor weight gain, flock non-uniformity, and diarrhea) and is associated with co-infections. Though unconfirmed, prior research suggests impaired nutrient acquisition or utilization. This study investigated how infection timing affects disease severity in turkey poults, hypothesizing that inoculation at early brooding (placement or 8 days of age) would lead to more severe outcomes relative to inoculation at 15 days of age or non-infected controls (NC). Poults were orally inoculated with a C. anatis field strain and performance recorded for 28 days. Ileal C. anatis concentrations and cecal tonsil concentrations of Escherichia coli, total coliforms, Clostridium perfringens, and Salmonella were measured at 28 days of age. Mortality was normalized with the Freeman-Tukey double arcsine transformation, and all parameters were assessed using a one-way ANOVA and Tukey-Kramer mean separation in JMP Pro 16. Mortality was significantly higher (p = 0.0007) with inoculation at placement and 8 days of age (up to 70 %) compared to those inoculated at 15 days of age (16 %) and the NC group (8 %). Initial weights were similar; however, poults inoculated at placement had significantly lower (p < 0.0001) final body weights (408.42 g/poult) compared to inoculation at 8 and 15 days of age (similar to 587 g/poult) and the NC group (797.51 g/poult). C. anatis and bacterial concentrations did not significantly differ. This study showed that C. anatis infection in early brooding leads to severe disease, highlighting the fragility of young poults susceptible to C. anatis infection.

Cochlosoma anatis is a flagellated protozoan parasite implicated in enteric disease (cochlosomiasis) of turkeys in commercial production. Research evaluating the effects of cochlosomiasis on turkey poults, where the disease is typically at its most severe, is lacking. Commonly observed cochlosomiasis symptoms include depressed weight gain, flock non-uniformity, lethargy, watery diarrhea, and co-infections. No commercially approved treatments or vaccines are available to combat cochlosomiasis. This study hypothesized that oral inoculation of C. anatis into turkey poults would lead to cochlosomiasis. Physiological and morphological analyses included blood chemistry, nutrient digestibility, and gut permeability. Disease effects were evaluated at 28 days of age in off-sex male turkey poults following inoculation with C. anatis at 14 days of age. Infection resulted in weight gain deficiencies; however, flock uniformity was not affected, contrary to field observations, which are likely related to the timing of infection relative to the bird’s age. Feed consumption was lower, and the feed conversion ratio was increased in infected cages. Amino acid digestibility was lower in infected birds, providing a potential partial explanation for the poor weight gain associated with cochlosomiasis. Blood chemistry analyses revealed metabolic alkalosis partially compensated for by respiratory carbon dioxide retention. Intestinal permeability was not significantly different between the groups. Overall, the disease effects observed in this study aligned mainly with traditional observations of cochlosomiasis. Flock non-uniformity may be the result of a complex of multiple issues that require exploration. The disease characterization presented here offers potential explanations, highlighting areas that suggest further research focus.

Cochlosoma anatis is a flagellated protozoan parasite classified in the Trichomonadidae family and is the causative agent of cochlosomiasis, an enteric disease of turkeys, waterfowl, and other wild birds. Cochlosomiasis symptoms largely consist of watery diarrhea, lethargic birds, depressed weight gain, and widespread flock morbidity causing flock nonuniformity. The known distribution of C. anatis is centered around areas of turkey production farms in the southeast United States, e.g., North Carolina, Missouri and Arkansas, but has been reported in other states and some other countries. Diagnosis is confirmed through examination of enteric mucosal scrapings using light microscopy. Following the withdrawal of approval of effective antiprotozoal medications for use in commercial animal production, cochlosomiasis has become a greater concern for commercial turkey industry professionals. Transmission of C. anatis occurs via the fecal-oral route, but the organism is fragile outside the host, suggesting the implication of a vector in the introduction of disease to susceptible farms. Research regarding C. anatis pathogenicity, transmission, and environmental involvement has been limited, creating a gap in cochlosomiasis knowledge. Future research is needed to further explore ways to prevent and treat cochlosomiasis, with needs centered on disease pathogenesis, transmission patterns, and prophylaxis and treatment methods.

The aim of the current study was to evaluate the influence of stressors on histomoniasis development and lateral transmission of Histomonas meleagridis. In the following experiments, half of the birds in each pen were inoculated with H. meleagridis to study disease transmission and progression. Birds were infected at 5 wk of age (experiments 1 and 3) or at 2 wk of age (experiment 2). Disease progression was evaluated by infection rate, mortality rate, and pathologic lesions in the ceca and liver. Reported results were applicable for directly infected birds as lateral transmission was not induced in these experiments. In experiment 1, the results showed high electrolyte, low-density diet (HE + LD), feed withdrawal (FW), caused higher infection rates and increase lesion scores in the liver and ceca compared to con. Experiment 2 further investigated the influence of low-density diet (LD) in conjunction with coccidiosis (LD + C) or feed withdrawal (LD + FW). All treatments had higher infection rates, mortality rates, ceca and liver scores compared to PC. In experiment 3, birds were fed diets containing naturally occurring aflatoxin at 0 ppb (AFLB1), 6.26 ppb (AFLB1 Low), or 19.82 ppb (AFLB1 High). No significant differences among treatments were observed. Though lateral transmission was not induced by theses stressors, the results of these experiments demonstrated that low-density diets, feed withdrawal and/or coccidial infection facilitated more severe histomoniasis infection.

Histomonas meleagridis, a protozoan parasite, induces blackhead disease (histomoniasis) in poultry. During hatching, chicks from lines divergently selected for high (HAS) and low (LAS) antibody responses to sheep red blood cells were divided into two groups, each of HAS and LAS, and placed in pens with wood shavings as litter. Feed and water were allowed ad libitum. Half of the chicks from each line had Limosilactobacillus reuteri (L. reuteri) inoculated to their drinking water. On day 18, all chicks were given a transcloacal inoculation of 100,000 H. meleagridis cells. Then, 10 days later, they were euthanized, followed by collection of tissues from the brain, cecal tonsil, ceca, liver, thymus, and spleen for qPCR analyses of cytokines involved in immunological development. Changes in cytokine expressions were most numerous in the cecal tonsil, ceca, and liver. In the absence of a functional medication for control of histomoniasis, L. reuteri and/or its secretory product, reuterin, might serve, in some genetic populations, as a means to reduce the impact of histomoniasis in chickens. The data demonstrate that L. reuteri treatment had tissue specificity between the two genetic lines, in which the effects were targeted primarily toward the cecal tonsil, ceca, and liver, which are the primary tissue targets of the parasite (H. meleagridis), as well as the thymus and spleen. However, interactions among main effects reflect that responses to inflammatory markers observed in tissues for one genetic line may not be observed in another.

A video received by faculty at North Carolina State University's Prestage Department of Poultry Science revealed a live parasite inside a chicken egg. The parasite was identified as an oviduct fluke (Prosthogonimus macrorchis), a trematode with a three-host life cycle: the primary host, a galliform bird, then an aquatic snail, and finally a dragonfly larva or adult consumed by the infected bird. The egg was from a "backyard flock" with access to a watercourse. No other instances of this parasite were seen in eggs from the flock. The presence of this parasite inside an egg suggests that the worms had migrated above the shell gland in the oviduct to be incorporated inside the egg. Currently, the occurrence of an oviduct fluke inside an egg in the United States is rare. Such parasites are not found in eggs from caged layers because those birds do not have access to watercourses. This case reinforces the view that parasites requiring intermediate hosts may become more common in birds reared under free-range conditions.

Coccidiosis is a high-prevalence disease that annually entails huge costs for the poultry industry. Control of coccidiosis in poultry production is based on the use of coccidiostats and vaccines. However, along with the problem of drug resistance, there is a concern about food safety and drug residues in poultry products. The objective of this study was to evaluate the effect of sodium bisulfate (SBS) in comparison with monensin (M) and their combination (SBSM) effects on controlling coccidiosis in broilers. In a randomized design, 300 chickens (Ross 308) were divided into 5 treatments and 4 replications (15 birds per replicate). All birds, except the negative control (NC), were orally inoculated with 4 Eimeria species on 14 D of age. Treatments included were as follows: NC, an unsupplemented basal diet, nonchallenged; positive control, a basal diet unsupplemented, challenged with Eimeria spp; a basal diet supplemented with 5 g/kg of SBS; a basal diet supplemented with 1 g/kg of M; and a basal diet supplemented with 5 g/kg SBS and 1 g/kg M (SBSM). Oocyst shedding per gram (OPG) of the faecal sample from each experimental unit was counted on 5 to 14 D after inoculation. Two chicks from each experimental unit were euthanized to investigate intestinal lesions on day 5 after inoculation. The NC birds showed the highest BW gain and the lowest feed conversion ratio. The birds in the SBSM group had improved feed consumption compared with theMgroup in the prechallenge period (P < 0.05). All supplemented treatments resulted in a significant decrease in OPG. The M and SBSM treatments showed more efficacy than the SBS group (P < 0.05) in reducing OPG. There was a significant reduction in cecal lesions owing to supplementation with SBS, but the effect of SBS in the upper part of the intestine was lower than the M and SBSM groups (P < 0.05). Based on the results of this study, SBS has protective effects against coccidiosis in ceca, and the combination of M and SBS (SBSM) did not show any further improvement effect compared with M alone on the control of coccidiosis.

A study was conducted to determine differences between Histomonas meleagridis–infected and control pullets based on disease signs, hen growth, and egg production and quality. Ross 708SF females were weighed and then placed in pens on the day of hatch (92 chicks/pen). At 25 D, 4 pens were infected with H. meleagridis in the cloaca, whereas 4 pens were control. At 5, 10, and 20 D after inoculation, 5 birds per pen (2 birds per pen at 20 D) were subjectively scored for blackhead disease. Birds were feed restricted based on BW and/or egg production. Individual BW were collected at 3, 5, 13, 15, 20, and 64 wk. Egg production was recorded at 24–63 wk. Egg quality was measured at 30, 34, 39, 42, and 56 wk and included shell and vitelline membrane strength, shell thickness, egg weight, and Haugh units. Hatchability was measured at 27, 37, and 60 wk and fertility at 27 and 37 wk. Treatment effects were determined by JMP Pro 14 using GLM with means separated using the Student t test (P ≤ 0.05). Cecal lesions were apparent on 5, 10, and 20 D and liver lesions on 10 and 20 D for the infected birds. The control had no histomoniasis lesions. Flock uniformity differed on wk 13 and 20 (P = 0.04; 0.04). Infected birds weighed less at 64 wk (P = 0.002). The onset of lay was not delayed. Infected birds produced more eggs during 1 period (P = 0.02). The infected birds produced heavier eggs at 30 wk (P = 0.04), eggs with a stronger and thicker shell at 42 wk (P = 0.05, 0.03), and eggs with a stronger vitelline membrane at 56 wk (P = 0.049). Hatchability and fertility did not differ (P > 0.05). H. meleagridis was observed in the infected birds' cecal samples at trial termination. This study indicates early infection with H. meleagridis has limited effects on pullet egg production and quality.

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.

Pathology and putative virulence factor expression of three Histomonas meleagridis isolates differing in geographic origin, cell passage number (56 or 100), or cell populations grown from a monoculture were compared. Turkey poults inoculated with the high cell passage number isolates or monoculture isolates varied in gross lesion severity and weight gain (P<0.0001). Screening of a published H. meleagridis cDNA library identified forty- eight cysteine proteinases (CP) and one superoxide dismutase (Fe-SOD) proposed to function in either tissue damage and/or invasion and oxidative defense. The Fe-SOD and eight CPs were analyzed using real time polymerase chain reaction. CP2, CP3, and CP8 showed significant differences in expression among the field isolates (P ≤ 0.05). The high passage isolates had decreased CP2, CP3 and CP4 expression when compared with their field isolate. CP7 did not differ between field isolates or the 56-passaged isolate. The Fe-SOD gene showed significant differences in expression among the various isolates. When exposing cultured H. meleagridis to air, Fe-SOD expression decreased rapidly during the first hour of air exposure but increased progressively through the next 3 h. This study provides information on gross pathology and virulence factors associated with various isolates of Histomonas meleagridis which can aid in determining the pathogenetic mechanisms used by this organism.

Sodium bisulfate (SB) was evaluated on its ability to improve broiler growth and intestinal structure with(out) a coccidia challenge. One thousand two hundred Cobb500 day-old males were randomly assigned within 4 experimental groups with a 2 × 2 factorial design, with (out) SB in the diet and with(out) a day 0 coccidia challenge using a 10× dose of a commercial vaccine. At day 7, oocysts per gram of feces were determined. At day 0, 14, 28, and 41, BW and feed consumption were measured. At day 21, 20 birds per treatment were subjectively scored for coccidia lesions, and jejunal histologic samples were collected for villi measurements. Twenty additional birds were given fluorescein isothiocyanate-dextran to determine gut permeability. At day 41, 10 birds per treatment had histologic samples collected. Statistical analysis was conducted in JMP Pro 14 using GLM procedure to compare disease state and diet. Means were separated using Dunnett's test (P ≤ 0.05) with the nonchallenged standard diet treatment that is considered the control. All parameters measured indicated an effect due to the coccidia inoculation. Therefore, effects of diet on (non)challenged treatments were determined using a Student t test (P ≤ 0.05). Limited differences due to diet were seen for the nonchallenged production data. Sodium bisulfate had a thinner villi base width (P = 0.04) on day 21 and greater villi height (P = 0.03), smaller base width (P = 0.04), thicker muscularis (P = 0.03), and lower crypt: height ratio (P = 0.01) on day 41. Challenged SB had similar gut permeability to the nonchallenged control (P = 0.94) on day 21. There was no difference in flock uniformity, feed intake, oocysts per gram of feces, or lesion scores between challenged treatments. Challenged SB had greater BW on day 14 (P < 0.0001), 28 (P < 0.0001), and 41 (P = 0.02). Feed conversion ratio from day 0 to 14 was also lower (P = 0.0002). Challenged SB had smaller crypts (P = 0.02) and therefore a smaller crypt: height ratio (P = 0.03) on day 21. Challenged control had a larger apical width (P = 0.03) and thicker muscularis (P = 0.04) on day 41. Overall, the addition of SB during coccidial enteropathy aided in BW, feed conversion ratio, and villi health with no observed effects on parasite cycling.

Nematodes are widespread and common in poultry. Disinfectants are used to reduce infection rates in poultry houses, but there is little documentation of their effectiveness. An in vitro assay was developed to test the efficacy of products to damage Heterakis gallinarum eggs, and nine disinfectants and chemicals commonly used in the poultry industry were tested. Embryonated eggs of H. gallinarum were pipetted into wells of plastic cell culture plates (250-300 eggs/well in water). Measured amounts of test articles were added to the suspensions for 2, 4, 6, or 24 hr. After exposure, eggs were washed with water and treated with trypan blue (1 ml of 0.4% solution, added to each well) at room temperature for 2 min. Eggshell integrity was determined microscopically by counting the number of eggs that were clear (intact) or that contained blue dye (compromised). As a test of embryo viability, five eggs per well from treatments containing compromised eggs were transferred to a Petri dish and hatched manually, using forceps to open the eggshell. Released larvae were then observed for signs of controlled movement. In a test of Clorox bleach (NaOCl), Green Klean, Decon7, Kem San, PLT, Virkon S, NaCl, dry limestone (CaCO3), and diesel fuel, only NaOCl (bleach) and Green Klean damaged the eggshell, and only 20,625 ppm of NaOCl rendered the larvae nonviable.

Blackhead disease, caused by the protozoan Histomonas meleagridis, is commonly found in layer pullets raised on the floor. We examined the effects of blackhead disease during the pullet-rearing period and on subsequent productivity during the first 8 wk of the laying cycle. Treatments were (1) uninfected controls and (2) H. meleagridis -infected pullets, with 4 replicate pens/treatment, 32 pullets/pen (Hy-LineW-36). Pullets in the challenge treatment were infected with H. meleagridis on day 18. Four birds/pen were necropsied on days 23 and 28 for lesion scores and day 176 for detection of H. meleagridis. Hens were moved to individual layer cages on day 120 and observed daily for feed consumption, date of first lay and egg production parameters. Pullets were positive for signs of blackhead disease in 83%–90% of infected birds necropsied on days 23 and 28, with average cecal lesion scores of 2.5 and 2.9. No liver lesions were observed. On day 176, 40% of infected birds were positive for H. meleagridis in the ceca. During the laying cycle, there were no significant differences (P ≤ 0.05) between treatments in terms of date of first lay, hen-day egg production, egg weight, feed conversion, egg mass/hen, or other reproduction measurements. These results showed that while there was no long-term effect of blackhead infection on layer productivity under laboratory conditions, H. meleagridis persisted in the flock, providing a reservoir for infection.

Chapter 28 Protozoal Infections Larry R. McDougald, Larry R. McDougaldSearch for more papers by this authorHector M. Cervantes, Hector M. CervantesSearch for more papers by this authorMark C. Jenkins, Mark C. JenkinsSearch for more papers by this authorMichael Hess, Michael HessSearch for more papers by this authorRobert Beckstead, Robert BecksteadSearch for more papers by this author Larry R. McDougald, Larry R. McDougaldSearch for more papers by this authorHector M. Cervantes, Hector M. CervantesSearch for more papers by this authorMark C. Jenkins, Mark C. JenkinsSearch for more papers by this authorMichael Hess, Michael HessSearch for more papers by this authorRobert Beckstead, Robert BecksteadSearch for more papers by this author Book Editor(s):David E. Swayne, David E. Swayne Laboratory Director Southeast Poultry Research Laboratory, U.S. National Poultry Research Center, Agricultural Research Service, U.S. Department of Agriculture, Athens, Georgia, USASearch for more papers by this authorMartine Boulianne, Martine Boulianne Professor Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Montreal, St-Hyacinthe, Quebec, CanadaSearch for more papers by this authorCatherine M. Logue, Catherine M. Logue Professor Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens, Georgia, USASearch for more papers by this authorLarry R. McDougald, Larry R. McDougald Professor Emeritus Department of Poultry Science, College of Agricultural and Environmental Sciences, University of Georgia, Athens, Georgia, USASearch for more papers by this authorVenugopal Nair, Venugopal Nair Head Viral Oncogenesis Group, The Pirbright Institute, Woking, Surrey, UKSearch for more papers by this authorDavid L. Suarez, David L. Suarez Research Leader Exotic and Emerging Avian Viral Diseases Research Unit, Southeast Poultry Research Laboratory, U.S. National Poultry Research Center, Agricultural Research Service, U.S. Department of Agriculture, Athens, Georgia, USASearch for more papers by this authorSjaak de Wit, Sjaak de Wit Poultry Veterinarian GD Animal Health, Deventer, The NetherlandsSearch for more papers by this authorTom Grimes, Tom Grimes Veterinary Consultant Grimes Consultancy, Paradise Point, Queensland, AustraliaSearch for more papers by this authorDeirdre Johnson, Deirdre Johnson Veterinarian Mountaire Farms, Inc., Millsboro, Delaware, USASearch for more papers by this authorMichelle Kromm, Michelle Kromm Director of Technical Services Jennie-O Turkey Store, Willmar, Minnesota, USASearch for more papers by this authorTeguh Yodiantara Prajitno, Teguh Yodiantara Prajitno Vice President Head of Poultry Health Services Head of Strategic Business Unit Animal Health Japfa Comfeed Indonesia, Jakarta, IndonesiaSearch for more papers by this authorIan Rubinoff, Ian Rubinoff Director of Sales and Technical Service, Europe Hy-Line International, Des Moines, Iowa, USASearch for more papers by this authorGuillermo Zavala, Guillermo Zavala Poultry Veterinarian Avian Health International, LLC, Flowery Branch, Georgia, USASearch for more papers by this author First published: 22 November 2019 https://doi.org/10.1002/9781119371199.ch28Citations: 5 AboutPDFPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShareShare a linkShare onFacebookTwitterLinked InRedditWechat Summary Protozoa found in poultry are categorized within several taxonomic groups. Of the numerous protozoans known from chickens, turkeys, and gamebirds, only a few flourish under commercial conditions. Parasites with short, direct life cycles, such as coccidiosis, are favored, whereas others involving intermediate hosts are normally not a problem in poultry. An exception is blackhead disease (histomoniasis), which has a complicated life cycle involving intermediate hosts, but takes advantage of reservoir hosts (chickens) and is able to spread easily from bird to bird within a flock of turkeys. Diagnostic tools for protozoan diseases in poultry have been developed, particularly with coccidia and Histomonas. Polymerase chain reaction-based tests are often used in research and may be useful to confirm species diagnosis. Knowing the route and source of protozoan infection is an important factor in developing biosecurity measures to decrease potential encounter with the bird and to break the parasites life cycle. Citing Literature Diseases of Poultry, 14th Edition RelatedInformation

Heterakis gallinarum is a widely distributed cecal nematode that parasitizes gallinaceous birds including chickens and turkeys. H. gallinarum infection poses a problem for the poultry industry as the nematode egg serves as a vector for the protozoan parasite, Histomonas meleagridis, the causative agent of histomonosis. The only means of detecting H. gallinarum in the environment is microscopic identification of the eggs in soil or feces; however, H. gallinarum eggs are often mistaken for those of Ascaridia galli. Three primer sets were designed from sequences cloned from the H. gallinarum genome to develop a diagnostic PCR. Each of these primer sets amplified a single product from H. gallinarum, but were unable to amplify DNA from H. meleagridis, Ascaridia galli, or Cestode sp. H. gallinarum DNA was amplified from Lumbricus sp. (earthworms) and Alphitobius diaperinus (darkling beetles), confirming that the earthworm acts as a paratenic host for H. gallinarum and suggesting that the darkling beetle may be a carrier for this nematode.

Heterakis gallinarum is a heavily prevalent poultry parasite that thrives in the ceca of various species of gallinaceous birds. It is a small roundworm, measuring between 4 and 15 mm long, in the family Heterakidae. Heterakis gallinarum has a direct life cycle not requiring an intermediate host to complete development, and it is generally believed that poultry raised at high density on litter are at greatest risk for accumulating large numbers of the nematode. This species typically only causes mild pathology that does not significantly affect bird performance. However, H. gallinarum is recognized as an economically important parasite by the poultry industry because its ovum serves as the vector for the protozoal parasite Histomonas meleagridis, the cause of histomonosis in poultry. Diagnosis of the nematode typically relies on fecal egg counts, which are prone to false negative diagnoses. Molecular tools are available for studying the nematode and diagnosing infected flocks. Treating and preventing H. gallinarum infection is made difficult due to the low efficacy of anthelmintics for eradicating H. gallinarum from infected birds and of disinfectants for destroying H. gallinarum ova on contaminated farms.

Blackhead disease is caused by Histomonas meleagridis, an anaerobic protozoan parasite, and results in mortality rates of up to 100% in turkeys and 30% in chickens. Outbreaks of blackhead disease are unpredictable, and the harvesting of H. meleagridis strains from the field would be a great resource for researchers to study its epidemiology. Therefore, the objective of this study was to develop a dry medium that would allow storage at ambient temperatures until needed. Fifty milliliters of horse serum was dried and then mixed with dry medium M199 with Hanks balanced salts (10.6 g), sodium bicarbonate (0.35 g), and rice powder (0.8 g). To test the ability of reconstituted medium to support growth of H. meleagridis, groups of 10 flasks containing 0.2 g of dry medium were stored for 24 hr at 25 and 60 C before testing. Other groups of flasks containing dry medium were stored at 25, 37, and 42 C for 1, 3, or 6 mo. At each test period, the flasks were reconstituted with 10 ml of water, inoculated with 100 000 H. meleagridis cells, and incubated at 40 C for 48 hr. Fresh liquid medium was used as a control. There were no differences in cell counts in medium stored at 25 or 60 C for 24 hr. After 1 mo, cell counts in reconstituted medium were about half that of fresh liquid medium after 48 hr of incubation. But after 3 and 6 mo, the cell counts were not significantly different in all groups (P < 0.05) after 72 hr of incubation. These results show that dried Dwyer medium can be stored at ambient temperatures for extended times and would be an effective tool for obtaining isolates of H. meleagridis from the field.

Histomonas meleagridis is a flagellate protozoan organism that can cause severe necrotizing typhlitis and hepatitis in gallinaceous birds. Peafowl (Pavo spp.) have been shown to be susceptible to histomoniasis in experimental settings, but there are few reports of natural histomoniasis in this species. A retrospective study of the archived cases at 2 veterinary diagnostic laboratories in the United States yielded 5 cases of peafowl with gross and histologic findings characteristic of histomoniasis. Lesions included bilateral, transmural fibrinonecrotic typhlitis and multifocal necrotizing hepatitis with associated trophozoites morphologically consistent with H. meleagridis. There was no evidence of Heterakis gallinarum infestation in the studied cases. DNA was extracted from formalin-fixed, paraffin-embedded liver and ceca from all 5 cases and was analyzed using multiple sets of primers with subsequent sequencing and genotyping. Four samples were positive for H. meleagridis, and 1 sample was positive for both H. meleagridis and Tetratrichomonas gallinarum. These results confirm that peafowl develop clinical disease similar to that described previously in other gallinaceous birds infected by H. meleagridis. The role of T. gallinarum remains unknown and further research is necessary to elucidate its role, if any, in the pathogenesis of the observed lesions.

Captive rearing and subsequent release of game birds, including northern bobwhites (Colinus virginianus), has become common in certain areas. In this practice, bobwhites are often raised in confinement to ‘flight ready’ and subsequently released for hunting. It is estimated that 30–40 million bobwhites are raised in captivity annually and some farms in the USA produce upwards of 1 million birds annually for this market. Raising game birds in these densities greatly facilitates the transmission of pathogenic organisms. Coccidiosis has been previously identified as an important disease in captive bobwhites and infection can lead to weight loss, diarrhea, poor feather growth, dehydration and, in severe cases, death. Eimeria lettyae, E. colini, and E. dispersa are the three described coccidia species from bobwhites. We investigated the prevalence and distribution of species of coccidia in captive bobwhite facilities throughout the United States. We collected litter or intestinal samples from 31captive bobwhite facilities originating from 13 states. Species-specific PCR primers were constructed against the internal transcribed spacer region 1 (ITS-1) of the ribosomal RNA gene of the various Eimeria spp. to aid in parasite detection and distinction. Primers were used to detect the specific Eimeria spp. in the collected samples. All 31 samples were positive for coccidia. Results of the primer survey disclosed E. lettyae, E. dispersa, and an unidentified Eimeria sp. in 20 (64.5%), 22 (72%), and 29 (93.5%) of the samples, respectively. Thirteen (41.9%) samples had 3 Eimeria spp. detected, 14 (45.2%) samples had 2 spp. detected, and 4 (12.9%) samples had 1 sp. detected. Flock age or geographical location was not associated with the presence of any particular Eimeria spp. To our knowledge, this is the first study of coccidia in captive bobwhites. Previous studies of Eimeria spp. in wild northern bobwhite are rare and disclosed variable prevalence rates ranging from 0 to 36%; no efforts were made to distinguish the coccidia species in these studies It would be helpful to use the species-specific primers constructed in this study to examine the prevalence and distribution of the Eimeria spp. in wild bobwhites from throughout their range to investigate the potential for captive-raised bobwhites to be a source of coccidiosis for wild bobwhites.

Anticoccidial sensitivity tests (ASTs) serve to determine the efficacy of anticoccidial drugs against Eimeria field isolates in a controlled laboratory setting. The most commonly measured parameters are body weight gain, feed conversion ratio, gross intestinal lesion scores, and mortality. Due to the difficulty in reliably scoring gross lesion scores of Eimeria maxima, microscopic analysis of intestinal scrapings (microscores) can be used in the field to indicate the presence of this particular Eimeria. The goal of this study was to determine the relationship between E. maxima microscores and broiler body weights and gross E. maxima lesion scores in three ASTs. Day-old broiler chicks were raised for 12 days on a standard corn-soy diet. On Day 12, chicks were placed in Petersime batteries and treatment diets were provided. There were six birds per pen, four pens per treatment, and 12 treatments, for a total of 288 chicks per AST. The treatments were as follows: 1) nonmedicated, noninfected; 2) nonmedicated, infected; 3) lasalocid, infected; 4) salinomycin, infected; 5) diclazuril, infected; 6) monensin, infected; 7) decoquinate, infected; 8) narasin nicarbazin, infected; 9) narasin, infected; 10) nicarbazin, infected; 11) robenidine, infected; and 12) zoalene, infected. On Day 14, chicks were challenged with an Eimeria field isolate by oral gavage. On Day 20, broilers were weighed, and gross lesion scores and microscores were classified from 0 to 4 depending on the severity of the gross lesion scores and E. maxima microscores. Data from three trials using different field isolates were statistically analyzed using a logarithmic regression model. There was no relationship (P = 0.1224) between microscores and body weight gain. There was a positive relationship between microscores and gross lesion scores (P = 0.004). However, there was also an interaction between isolate and treatment (P < 0.0001). Lastly, the interaction between isolate and gross lesion scores (P = 0.0041) demonstrates that the significance of the relationship between microscores and gross lesion scores may be dependent on pathogenicity of the challenge Eimeria or the amount of E. maxima in the inoculum.