Experimental caprine coccidiosis caused by Eimeria arloingi: Morphopathologic and electron microscopic studies

The progressive morphohistopathologic changes, distribution pattern of lesions and ultrastructural characteristics in Eimeria arloingi infection were investigated in experimentally infected kids. The 18 newborn animals allocated to 3 equal groups. Two of groups, A, B were inoculated with a single dose of 1 × 10³ and1 × 10⁵ sporulated oocysts of E. arloingi, respectively. At 7, 14, 21, 28, 35, and 42 days postinoculation (DPI), 1 kid from each group was necropsied for pathologic and ultrastructural studies. Progressive lesions were present at 21, 28, 35 and 42 DPI in the jejunum, ileum, cecum with fewer in the duodenum and proximal colon. The oocysts shedding begin between 16 to 18 DPI. Grossly, minimal changes were observed at 21 DPI as few whitish plaques or nodules and advanced lesions at 42 DPI as pseudoadenomatous pattern in the mucosa and a cerebriform pattern on the serosal surface of jejunum and ileum. Early histopathologic lesions due to schizogony phase were including presence of intracytoplasmic developmental stages of the parasite such as trophozoites, immature to mature schizonts and mild infiltration of inflammatory cells. In late lesions due to various stages of gametogony, the histological pattern was mainly remarkable hyperplasia of the villi and crypts epithelial cells, eventually developed into papillary projections of reactive epithelium. The mesenteric lymph nodes showed a few numbers of large schizonts in the cortical lacteals. This study showed E. arloingi as a highly pathogenic species for kids, the incubation period was 16–18 days and the main target organ was jejunum with characteristic morphohistopathologic lesions.     

Lesions in goats fed Toxoplasma gondii oocysts

Tissues of 20 2-3-month-old kids and 17 2-4-year old does fed Toxoplasma gondii oocysts were examined histologically. Kids were necropsied between 3 and 50 days post-inoculation (DPI) and does were necropsied between 10 and 422 DPI. Lesions were seen in tissues of all young goats fed 1000-100,000 oocysts and consisted of acute enteritis, necrosis of mesenteric lymph nodes and encephalomyelitis. Lesions were not seen in does fed 10-10,000 oocysts.     

Progressive immunopathological changes during early stages of experimental infection of goats with Mycobacterium avium subspecies paratuberculosis

A dose of 10(10) Mycobacterium avium subspecies paratuberculosis was administered orally on seven occasions to produce experimental paratuberculosis infection in 10 5-8-week-old goat kids. Bacteriological, immunological, and histopathological changes, their relationships, and the efficacy of the commonly used diagnostic methods were studied during the progressive disease up to 270 days postinfection (DPI). Significant lymphocyte proliferative responses in the peripheral blood of five goats were detected as early as 60 DPI. A lymphoproliferative test was also performed on lymphocytes purified from different compartments of the guts of five infected and five control goats. Significant proliferative responses were observed in lymphocytes of jejunal compartments of all five goats, of which four had also significant lymphocyte proliferation in the blood. The ileal lymphocytes from two goats, one each at 120 and 270 DPI, had significant proliferation. The histological lesions were mainly observed in the gut-associated lymphoid tissues of the ileocecal valve, the ileum, and the terminal jejunum. Acid-fast bacilli were demonstrated in the lesions of two goats at 60 and 210 DPI. Bacterial culture showed poor sensitivity, detecting positive results for only one goat in the fecal and tissue samples at 210 DPI, whereas polymerase chain reaction (PCR) detected one goat in fecal sample at 210 DPI and two goats in tissue samples at 60 and 210 DPIs, respectively. Enzyme-linked immunosorbent assay and agar gel immunodiffusion test were found to be 100% sensitive from 180 and 210 DPI onwards, respectively.     

Sequential Clinical,Haematological, Biochemical,Immunopathological and Histopathological Alterations in Buffalo Calves (Bubalus bubalis) Intravenously Infected with Absidia corymbifera

Absidiosis was produced experimentally in 18 buffalo calves by intravenous inoculation of spores of Absidia corymbifera. Infected animals exhibited dullness, depression, partial anorexia and an initial pyrexia and coughing during the first week and two animals died on each of 9, 13 and 16 days post infection (DPI). The haemoglobin concentration and total erythrocyte count showed no appreciable change from their basal values at any stage of the experiment. However, the erythrocyte sedimentation rate and total leukocyte count increased significantly in the infected animals. The differential leukocyte count revealed a relative neutrophilia from 5 to 20 DPI. There was a significant increase in the serum total proteins, blood urea nitrogen, creatinine, alanine aminotransferase, aspartate aminotransferase, serum alkaline phosphatase, total immunoglobulins and circulating immune complexes in the infected animals as compared to the controls. In the sera of the infected animals, specific Absidia corymbifera IgM and IgG antibodies were detected from 3 DPI to 6 DPI respectively by Dot-EIA. Type I and type III skin hypersensitivity were detected from 10 DPI and type IV hypersensitivity from 15 DPI onwards. The gross and microscopic pathological lesions were seen mainly in the lungs, in all except one of the affected animals. This animal died 9 DPI and mycotic granulomas were also seen in its heart and kidneys. The microscopic lesions in the lung took the form of well-developed granulomas.     

Histochemical alterations in the duodenum of goats experimentally infected with Paramphistomum cervi

Certain histochemical alterations in the different tunics of duodenum in kids were studied 20, 40, 60 and 80 days post-infection (DPI) with Paramphistomum cervi and the results compared with those of uninfected kids. There was a general reduction of polysaccharide complex substances and glycogen at 20 DPI. A marked decrease in polysaccharide complex substances and glycogen at 20 DPI. A marked decrease in polysaccharide complex substances and glycogen was especially discernible in the Brunner's gland and muscularis mucosa 20 DPI. Thereafter, these substances gradually increased and at 80 DPI this decrease was fully replenished. A slight reduction in mucin and protein content of the infected duodenal goblet cells was noticed at 20 DPI. It is suggested that juvenile P. cervi utilize host-tissue polysaccharide complex substances and glycogen for their growth and development during duodenal migration.     

Lesions in transplacentally induced toxoplasmosis in goats

Fifteen pregnant does were inoculated orally with 10 to 1,000 oocysts of the GT-1 strain of Toxoplasma gondii. Two does were euthanatized on postinoculation days (PID) 10 and 14, and the remaining 13 does aborted or delivered kids transplacentally infected with T gondii on PID 9 to 65. Tissues of all 34 fetuses or kids from the inoculated does were examined for gross and microscopic lesions. Placental necrosis and encephalomyelitis were the most prominent lesions. Toxoplasma gondii invaded the fetal placenta between PID 9 and 11 and invaded fetal tissues 2 to 3 days later. Necrosis attributed to toxoplasmosis, confined to cotyledons, was found in all placentas examined on PID 18. Encephalomyelitis was found in most fetuses or kids between PID 30 and 65. Lesions in fetal tissues were sparse. Because T gondii is rarely found in lesions, histologic examination of the fetal tissues is not likely to help in diagnosing every case of transplacentally induced toxoplasmosis in goats.     

Recovery of Mycoplasma agalactiae from the ears of goats experimentally infected by the intramammary route

The role of inapparent carriers of Mycoplasma agalactiae and the strategies used to colonise the external ear canal in goats remain unclear. This study examined the ability of M. agalactiae to colonise the ears of goats infected experimentally by the intramammary route. The right mammary glands of 15 lactating goats were inoculated with 10(10) colony forming units (cfu) of M. agalactiae. The goats were randomly assigned to three groups of five animals each and sampled at slaughter at 5, 15 or 45 days post-infection (dpi). A further four goats served as uninfected controls. Right and left ear swabs were collected for detection of M. agalactiae by culture before and after sacrifice. M. agalactiae was detected in 19/20 (95%) ear swabs from goats sampled at 15 and 45dpi, whereas all ear swabs collected before inoculation, ear swabs collected from the group sampled at 5dpi and ear swabs from control goats at the time of sacrifice were negative for M. agalactiae. Blood samples collected at 6, 12, 24, 48 and 72h post-infection for detection of M. agalactiae by culture were also negative. There were differences in the antigenic profiles of isolates recovered from the ears compared to the 7MAG strain used to inoculate the animals and most isolates from the mammary gland, milk and supramammary lymph nodes.     

Application of different methods for the diagnosis of experimental paratuberculosis in goats

The diagnosis of subclinical paratuberculosis is still considered a major problem worldwide. As part of investigating diagnostic strategies for the paratuberculosis infection, sequential results of various diagnostic methods in a progressive experimental infection in goats were evaluated. Twenty-three goat kids were divided into three groups: the infected, contact and control, comprising 10, five and eight goats respectively. Animals of the infected group were orally inoculated on seven occasions with 5 ml of inoculum containing 2 x 10(9)Mycobacterium avium ssp. paratuberculosis per ml. Lymphoycte proliferation test using johnin PPD detected paratuberculosis infection from 60 days post-infection (DPI) onwards. The johnin PPD was found to be a better antigen for the proliferative assays as compared with the sonicated antigen. The faecal smear examination with acid-fast staining detected more goats as positive than bacterial culture and polymerase chain reaction (PCR). Lipoarabinomannan enzyme-linked immunosorbent assay (ELISA) started detecting infected goats from 150 DPI onwards followed by indirect ELISA and agar gel immunodiffusion from 180 DPI onwards. Histological examination was confirmatory and detected five infected goats as positive.     

Intestinal lesions caused by a strain of Chlamydia suis in weanling pigs infected at 21 days of age.

The objective of this study was to determine whether a strain of Chlamydia suis shown previously to be an intestinal pathogen in gnotobiotic piglets could cause diarrhea and intestinal lesions in young weanling pigs. Pigs from 2 sows were randomly assigned to 2 groups. Group 1 included 13 pigs that were weaned at 24 hours of age and then housed in isolator units and fed milk replacer and unmedicated starter ration. Group 2 included 8 pigs that nursed their respective sows, consumed unmedicated starter ration, and were weaned at 21 days of age. Ten pigs in group 1 and 6 pigs in group 2 were inoculated orally with 4 x 108 inclusion-forming units of C. suis strain R27 at 21 days of age. Control pigs were inoculated with sham inoculum. The pigs were necropsied 5-14 days postinoculation (DPI). None of the Chlamydia-infected pigs developed diarrhea. Villus atrophy was seen histologically in sections of ileum from Chlamydia-infected pigs in both groups 5 and 7 days DPI. Lymphangitis and multiple lymphohistiocytic and neutrophilic aggregates were seen in the submucosa, tunica muscularis, and serosa of the distal jejunum, ileum, and colon from Chlamydia-infected pigs in both groups 5-14 DPI. Immunostaining of sections of distal jejunum, ileum, and colon from infected pigs revealed chlamydial antigen in intestinal epithelium and in foci of lymphangitis/inflammation. The results indicated that C. suis strain R27 can cause intestinal lesions in young weanling pigs, and the lesions are similar to those seen in gnotobiotic piglets. The results also indicated that strain R27 causes asymptomatic intestinal infections in young weanling pigs, at least under the conditions of this study.     

Experimental and serologic observations on avian pneumovirus (APV/turkey/Colorado/97) infection in turkeys

An avian pneumovirus (APV) was isolated from commercial turkeys in Colorado (APV/Colorado) showing clinical signs of a respiratory disease. The results of virus neutralization and indirect fluorescent antibody tests showed that the APV/Colorado was partially related to APV subgroup A but was unrelated to APV subgroup B. Turkeys experimentally inoculated with the APV/Colorado were observed for signs, lesions, seroconversion, and virus shedding. Thirty-six 7-wk-old turkeys were distributed into three groups. Eighteen turkeys were inoculated oculonasally with APV/Colorado, six were placed in contact at 1 day postinoculation (DPI), and 12 served as noninoculated controls. Tracheal swabs and blood samples were collected at 3, 5, 7, 10, 14, and 21 DPI. Tissues were collected from three inoculated and two control turkeys on aforementioned days for pathologic examination and APV isolation. Inoculated turkeys developed respiratory disease, yielded APV at 3, 5, and 7 DPI, and seroconverted at 10 DPI. Contact turkeys yielded APV at 7 and 10 DPI. No gross lesions were observed in the turbinates, infraorbital sinuses, and trachea. However, microscopic examination revealed acute rhinitis, sinusitis, and tracheitis manifested by congestion, edema, lymphocytic and heterophilic infiltration, and loss of ciliated epithelia. The inflammatory lesions were seen at 3 DPI and became extensive at 5 and 7 DPI. Active regenerative changes in the epithelia were seen at 10 and 14 DPI. Serologic survey for the presence of antibodies in commercial turkeys (24,504 sera from 18 states) and chickens (3,517 sera from 12 states) to APV/Colorado showed seropositive turkeys in Minnesota, North Dakota, and South Dakota and no seropositive chickens. This report is the first on the isolation of an APV and APV infection in the United States.     

Chronologic localization of mycoplasma hyopneumoniae in experimentally infected pigs

The chronologic localization of Mycoplasma hyopneumoniae was examined by in situ hybridization in experimentally infected pigs for a period of 35 days after intratracheal inoculation. M. hyopneumoniae DNA was detected in bronchial and bronchiolar epithelial cells from infected pigs at 7, 14, 21, and 28 days postinoculation (DPI) and in alveolar and interstitial macrophages and type I pneumocytes from infected pigs at 14, 21, 28, and 35 DPI. Strong hybridization signals for M. hyopneumoniae were detected mainly at the luminal surface of bronchial and bronchiolar lining epithelial cells. When a hybridization signal was detected at the luminal surface of bronchial and bronchiolar lining epithelial cells, a given bronchus or bronchiole also exhibited peribronchiolar lymphoid cuffing. These observations suggested that the presence of M. hyopneumoniae in different tissues could be due to a difference in the duration of the infection.     

Blackhead disease in turkeys: direct transmission of Histomonas meleagridis from bird to bird in a laboratory model

The spread of Histomonas meleagridis infections through groups of turkeys in the absence of the cecal worm vector (Heterakis gallinarum) was studied in a battery cage model. Battery-reared poults were exposed at 2 wk of age by commingling with infected birds into cages that had the floor lined with paper. One treatment received no exposure, whereas other birds were commingled with two, three, or four birds/cage (25%, 37.5%, or 50%) inoculated per cloaca with cultured H. meleagridis (200,000/bird). Inoculated birds died at 7-13 days postinoculation (DPI) showing typical liver and cecal lesions of histomoniasis. By 14 DPI, 87.5% of the directly inoculated birds died or had severe lesions of histomoniasis. Turkeys commingled with two, three, or four infected birds became infected at the rate of 72%, 80%, or 75%, respectively. In another experiment, two birds/cage (25%) were inoculated with Histomonas from culture and allowed to commingle with other birds for 1, 2, 3, or 4 days. Two of 12 (16.7%) birds had minor cecal lesions after contact with inoculated birds for 1 day, but 87.5%-100% became infected if inoculated birds remained in the cage for 2-4 days. Contemporaneous inoculation with cecal coccidia (Eimeria adenoeides) as a predisposing factor in blackhead infections was studied using the model. Turkey poults directly inoculated with Histomonas were allowed to commingle for 5 days with uninoculated birds that had received inoculation with 0, 10(3), or 10(4) sporulated oocysts. The coccidian infection appeared to interfere with transmission of blackhead infection by 7 DPI, as suggested by lessened severity of cecal lesions and a lower percentage of infected birds. These studies confirm that histomoniasis is transmitted readily from directly exposed young turkeys to others in the absence of the cecal worm vector, and that this phenomenon can be reproduced in battery cages as an experimental model.     

Goat flea (order Siphonaptera) as a possible vector for the transmission of caprine mycoplasmal polyarthritis with septicaemia

Goat fleas of the order Siphonaptera were collected from the body surfaces of naturally infected polyarthritic goat kids with septicaemia. These fleas and the infected kids were positive for Mycoplasma mycoides subsp. mycoides (large colony type) identified by cultural, morphological, biochemical and growth inhibition tests. Blood from the infected fleas contained 1 × 10²−1 × 10⁵ viable subsp. mycoides ml⁻¹. Experimental transmission of the disease to other kids was established by placing 120 infected fleas on each kid's body surfaces. All experimentally infected kids developed characteristic clinical signs and showed leucopenia with neutropenia, an increased amount of fibrinogen and mortality with lesions of suppurative polyarthritis associated with septicaemia. The organisms were also recovered in high numbers from heart blood, body fluids, and infected organs and joints. The results suggested that fleas of the order Siphonaptera acted as vectors for the transmission of the spontaneous disease in kids.     

Induction of mycoplasmosis in goat kids by oral inoculation with Mycoplasma mycoides subspecies mycoides

A one-time, orally administered dose of greater than or equal to 1 X 10(6) colony-forming units of Mycoplasma mycoides subspecies mycoides was sufficient to induce clinical mycoplasmosis (n = 37) terminating in fatal mycoplasmemia in 73% (37 of 51) of the clinically affected kids. The pathogen was isolated from the blood samples as early as 24 hours after oral inoculation; hot, swollen joints frequently were evident by 4 or 5 days after exposure. Pyrexia (to 42.3 C) was detected in about 95% (35 of 37) clinically affected kids, although about 5% (2 of 35) died peracutely without fever or other premonitory signs. At necropsy, the cardinal lesions were a fibrinopurulent polyarthritis and red, patchy to diffuse areas of consolidation in 1 or more lung lobes. At death, usually within 4 to 16 days after oral inoculation, the concentration of M mycoides subspecies mycoides in the blood was 1 X 10(6) to 1 X 10(7) colony-forming units/ml. Histologically, the kids had diffuse fluid leakage into pulmonary alveoli and to a lesser extent into small vessels of various other organs. Fibrinocellular thrombi of terminal occurrence were occasionally present in various organs. The meningeal, pleural, and peritoneal surfaces had vascular leakage and a minimal perivascular accumulation of leukocytes. The disease was contagious. Of 14 noninoculated control kids in close confinement with affected kids, 8 (57%) developed mycoplasmosis in 7 to 15 days and died of mycoplasmemia. The remaining 5 noninoculated kids remained healthy, as did noninoculated kids that were kept isolated from affected kids.     

An outbreak of neurofilariosis in young goats

An outbreak of cerebrospinal microfilariosis in 23 kids, representing a whole kid crop, occurred on a farm in Qassim, Central Saudi Arabia. The morbidity and mortality rates were 100%. Clinical signs, in kids from the fourth to sixth week of age, manifested as weakness in the hind legs followed by incoordination of movement and feet dragging. Seven to ten days later, the hind legs were completely paralyzed, and within a week the front legs were also paralyzed. The animals continued to feed, drink and suckle normally, and were in good health. Paralyzed kids were helped to suckle by the farm attendants. With time, loss of control of motor functions occurred and the ability to hold the head in upright position was lost, ability to eat or drink was lost. The infected kids health deteriorated and they eventually died. Two dead kids were postmortemed. With the exception of a meningeal haemorrhage seen in one kid, no obvious macroscopic lesions were seen in any of the autopsied animals. Histopathological examination of the spinal cord and brain showed different microfilaria of different sizes between the meninges and the nervous tissue. This indicated that development of the parasite occurred in the nervous tissue. The larger microfilaria contained black deposits, most likely to be digested blood, in their guts. It is likely that the parasites are haematophagus. The main lesions caused by the microfilaria were migratory traumatic haemorrhagic tracts in the spinal cord and the brain. These sometimes contained parts of the microfilariae. In the brain malacic lesions and sporadic aggregates of lymphocytes were also noted.     

An experimental study on the pathogenicity of the caprine herpesvirus type 1 (CHV-1).

The pathogenicity of caprine herpesvirus type-1 (CHV-1) in goat kids and lambs was studied. Two experiments were carried out. In the first, two Saanen goat kids and four lambs of a local breed were infected intravenously with the Swiss strain E/CH of the virus. Clinical reaction was severe in the kids, but it was very mild in the lambs. Virus was excreted from the kids in higher titres and longer than in lambs. Virus was also isolated from tissue specimens but only from a kid that died on post inoculation day 4 (PID 4). The gross- and histopathological lesions were more severe in kids. In the second experiment only lambs were used. They were dexamethasone treated and then virus inoculated. A very mild infection developed. The lambs did not shed the virus, neither the virus was isolated from their tissues collected at necropsy. Nevertheless histopathological lesions were seen. In both experiments the animals seroconverted on PID 10.     

The effect of colostrum-derived antibody on neo-natal transmission of caprine arthritis-encephalitis virus infection

Two groups of 6 newborn goat kids were artificially fed colostrum containing antibody to caprine arthritis-encephalitis (CAE) virus, obtained from clinically affected does. Kids in group A were fed the colostrum from birth until 7 days of age, while kids in group B were fed colostrum from 1 to 3 days after birth for 7 days. Kids were fed cow's milk at all other times. Serum antibody resulting from the consumption of colostrum, detected by agar gel immunodiffusion (AGID) tests, lasted for up to 8 weeks in group A, but none was detected in group B. Four kids from each group became infected with CAE virus as demonstrated by the emergence of active immunity and by virus isolation procedures. It appeared that uptake of colostral antibody by group A did not prevent viral transmission, interfere with development of active immunity, or modify the outcome of the CAE virus infection.     

Pathogenesis of intestinal cryptosporidiosis in conventional and gnotobiotic piglets.

The pathogenesis of intestinal cryptosporidiosis was studied in 52 conventionally reared and 20 gnotobiotically reared piglets by inoculation with different doses of Cryptosporidium parvum oocysts. The prepatent period of C. parvum in both groups of animals were variable, depending on the number of oocysts administered. The patent period of C. parvum in conventionally reared piglets was 8 or 9 days; in gnotobiotic piglets cryptosporidia were found in feces until Day post infection (DPI) 16, when the last piglet was necropsied. Cryptosporidiosis in conventionally reared piglets is a self-limited diarrheal disease associated with morphological changes within the intestine. The most severe lesion was seen in the posterior jejunum and ileum from DPI 3 to DPI 7, and consisted of villous atrophy, crypt hyperplasia and inflammatory infiltration in the lamina propria. In gnotobiotic piglets cryptosporidia induced severe enterocolitis which occurred at least until DPI 16. The characteristics of enteric lesions were similar to those found in conventionally reared piglets. Intestinal cryptosporidiosis in both groups of animals shifted in the course of infection in the caudal direction and terminated in the large intestine. Examination by scanning electron microscope showed that infected absorptive cells had thicker and longer microvilli than those on non-infected cells; neighboring non-infected cells were hypertrophic, bulbously protuberant with minute microvilli with no distinct intercellular borders. Numerous cryptosporidia in the heterotopic glandular epithelium in the submucosa of cecum and colon on DPI 9 and 10 were found. No differences in the location and degree of cryptosporidial infection between colostrum-fed and colostrum-deprived conventionally reared piglets were found. Sow's colostrum does not appear to protect piglets from C. parvum infection. The role of intestinal microflora in the pathogenesis of cryptosporidiosis in piglets is discussed.     

Toxoplasma gondii-induced abortion in dairy goats

Three of 8 goats on a Maryland farm aborted or had dystocia associated with toxoplasmosis during the winter of 1984. Doe 1 aborted a decomposed fetus 30 days before term. Modified agglutination test (MAT) antibody titers against Toxoplasma gondii were found in pleural fluid of the fetus (1:1,024) and in serum of doe (1:4,096 at 31 days after abortion). Doe 2 aborted a fetus 5 days before term; MAT antibody was found in the pleural fluid of the fetus (1:16,384) and in the doe's serum (1:4,096 on the day of abortion). Placenta from both does had foci of necrosis characteristic of toxoplasmosis, and T gondii was identified in lesions. Doe 3 had dystocia 7 days before term and a partially decomposed fetus was delivered by cesarian section; MAT was found in pleural fluid of the fetus (1:1,024) and in serum from the doe (1:4,096 on the day of abortion). Focal gliosis and calcification were seen in brain specimens from 2 of the 3 fetuses. None of the does produced milk after abortion. Two other does (No. 4 and 5) delivered apparently healthy kids transplacentally infected with T gondii; MAT in serum of both does was 1:4,096. Doe 4 delivered 3 kids; MAT titer in a serum from each kid 38 days after birth was 1:16,384. Doe 5 delivered 1 kid with a serum MAT titer of 1:1,024 at 38 days after birth. The 3 remaining does had MAT titers of 1:256, 1:16, and 1:16, and all delivered healthy kids. Epizootiologic evidence suggested that the does acquired T gondii infection from oocysts passed in feces of domestic cats on the farm. The MAT titers of 4 cats on the farm were 1:65,356; 1:1,024; 1:16; and 1:1,024.