Comparison of the complement fixation test and competitive ELISA for serodiagnosis of Anaplasma marginale infection in experimentally infected steers

OBJECTIVE: To compare sensitivity of a complement fixation (CF) test and competitive ELISA (cELISA) for detection of Anaplasma marginale in experimentally infected steers. ANIMALS: 40 crossbred (Angus-Simmental) steers. PROCEDURES: Steers were inoculated with 2.6 x 10(9) A marginale-infected erythrocytes (day 0). Blood samples were collected on days 9, 13, 20, 28, 34, 41, 61, 96, 126, and 156 days after inoculation. The percentage of parasitized erythrocytes (PPE) was determined by microscopic examination of stained blood films, and sera were evaluated with the CF test and cELISA by use of USDA-approved methods. Sensitivity and agreement (kappa statistic) between the 2 methods were determined. Persistent infections were confirmed by inoculation of blood obtained from infected steers into susceptible, splenectomized calves. RESULTS: 9 days after inoculation, sensitivity of the cELISA was 47.5%, whereas the CF test failed to identify seropositive steers. After day 13, sensitivity of the cELISA and CF test was 100% and 20%, respectively. During peak parasitemia (day 20), sensitivity of the cELISA and CF test was 100%. Thereafter, sensitivity of the CF test fluctuated between 7.5% and 37.5%, whereas sensitivity of the cELISA remained at 100%. Overall sensitivity of the cELISA and CF test was 94.8% and 26.5%, respectively (kappa statistic, 0.039). CONCLUSIONS AND CLINICAL RELEVANCE: The cELISA had superior sensitivity for serologic detection of A marginale.The CF test and cELISA each had a high percentage of false-negative results during the prepatent period. These findings are relevant for export certification and anaplasmosis prevention or eradication programs.     

Temporomandibular joint ankylosis in cats and dogs. A report of 10 cases.

Temporomandibular joint (TMJ) ankylosis is characterized by difficulty or inability to open the mouth. The ankylosis may be articular ('true') or extra-articular ('false'). Clinical signs, radiographic studies, treatment and follow-up are presented in a retrospective study involving five cats and five dogs. The findings were compared with TMJ ankylosis in humans. CT imaging with three-dimensional reconstruction proved to be of great value in determining the extent of the abnormalities and helped with preoperative planning. Articular TMJ ankylosis occurred in six animals and extra-articular TMJ ankylosis was found in the other four cases. In three cats and in three dogs, the TMJ ankylosis was trauma related; the remaining patients were diagnosed with a tumour. Resection of ankylosing tissue in false ankylosis or gap arthroplasty in true ankylosis was successful in all of the trauma induced cases. In the two cats, with tumour related ankylosis, the ankylosis was caused by an osteoma and resection had a good prognosis, whereas the two dogs had to be euthanatized.     

Postenucleation orbital sialocele in a dog associated with prior parotid duct transposition

An orbital sialocele developed in a dog following enucleation for protracted glaucoma. This eye had historically been treated for keratoconjunctivitis sicca by parotid duct transposition approximately 5 years previously, and the duct was ligated distally at the time of enucleation. One month following enucleation, the dog presented with a fluctuant conical-shaped swelling ventrolateral to the orbital socket. Surgical exploration revealed a dilated, fibrotic distal portion of the previously transposed parotid duct, and saliva, within the enucleated orbit. The distal portion of the duct and saliva-containing tissues from within the orbit were excised. The remaining proximal normal portion of the parotid duct was re-routed into the oral cavity. Clinicopathologic and histologic examination of the excised orbital contents and dilated portion of duct revealed a sterile sample of saliva and moderate chronic periductal fibrosis. At a 6-month re-evaluation there was no evidence of recurrence of the sialocele, and the parotid duct was functional.     

Regional seroprevalence of bluetongue virus in cattle in Illinois and western Indiana

OBJECTIVE: To estimate seroprevalence of bluetongue virus (BTV) and the geographic distribution of seropositive cattle herds in Illinois and western Indiana. SAMPLE POPULATION: 10,585 serum samples obtained from cattle in 60 herds during 3 transmission seasons (2000 through 2002). PROCEDURES: In a longitudinal study, serum samples were tested for BTV antibodies by use of a competitive ELISA. Four geographic zones were created by use of mean minimum January temperature. A multivariable mixed-effects logistic regression model with a random effect for herd was used to estimate seropositive risk for zone, age of cattle, herd type, and transmission season. RESULTS: Overall, BTV antibodies were detected in 156 (1.5%) samples. Estimated seroprevalence in 2000, 2001, and 2002 was 1.49%, 0.97%, and 2.18%, respectively. Risk of being seropositive for BTV was associated with geographic zone and age. Seroprevalence increased progressively from northern to southern zones, with no evidence of BTV infection in the northernmost zone. In the southernmost zone, annual seroprevalence ranged from 8.65% to 11.00%. Adult cattle were 2.35 times as likely as juvenile cattle to be seropositive. CONCLUSIONS AND CLINICAL RELEVANCE: Overall seroprevalence was lower than has been reported for Illinois cattle. Bluetongue virus antibodies were distributed heterogeneously in this region. Only in the southernmost zone was seroprevalence consistently > 2%. Regionalization of BTV risk based on state borders does not account for such variability. Serologic data could be combined with landscape, climate, and vector data to develop predictive models of BTV risk within transitional regions of the United States.     

Efficacy of bovine herpesvirus-1 inactivated vaccine against abortion and stillbirth in pregnant heifers

OBJECTIVE: To evaluate the efficacy of an inactivated bovine herpesvirus-1 (BHV-1) vaccine to protect against BHV-1 challenge-induced abortion and stillbirth. DESIGN: Prospective study. ANIMALS: 35 beef heifers. PROCEDURES: Before breeding, heifers were vaccinated with a commercially available BHV-1 inactivated vaccine SC or IM. The estrous cycle was then synchronized, and heifers were artificially inseminated 30 to 60 days after vaccination. Heifers (n = 21) were challenge inoculated IV at approximately 180 days of gestation with virulent BHV-1. Fourteen control heifers were not vaccinated. Clinical signs of BHV-1 infection were monitored for 10 days following challenge; serologic status and occurrence of abortion or stillbirth were evaluated until time of calving. RESULTS: 18 of 21 (85.7%) heifers that received vaccine were protected from abortion following challenge, whereas all 14 control heifers aborted. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that an inactivated BHV-1 vaccine can protect against abortion resulting from a substantial challenge infection, with efficacy similar to that of modified-live BHV-1 vaccines.     

Comparison of anthelmintic activity of pyrantel, praziquantel, and nitazoxanide against Anoplocephala perfoliata in Horses

Three anthelmintics were compared for efficacy in reducing the egg production of Anoplocephala perfoliata in a herd of central Texas horses. Two trials were run, 1 in mares and the other in weanlings that were diagnosed as being infected with Anoplocephala by recovery of eggs in 5 g of feces with sugar centrifugation. Each animal was evaluated twice before treatment and again twice following treatment (at weeks 2 and 4 after treatment). The criteria for infection were the recovery of eggs on at least 1 occasion before treatment and the finding of eggs on 1 day following treatment. The mares were treated 1 time with either pyrantel pamoate at 13.2 mg/kg, nitazoxanide at 100 mg/kg, praziquantel at 1.23 mg/kg or remained as untreated controls. The weanlings were treated with pyrantel at 13.7 mg/kg nitazoxanide at 100 mg/kg or remained as untreated controls. The percentage reduction of patient infection in mares after treatment with pyrantel was 83%, with nitazoxanide was 78%, and with praziquantel was 83% and in controls was 17%. There was a 75% reduction of patient weanlings treated with pyrantel or nitazoxanide and a 17% reduction in untreated controls. The reduction of infection in all horses treated with any drug was significantly different from controls. All of the drugs were somewhat effective in the control of Anoplocephala, and there were no differences among the drugs in their effectiveness.

Introduction

Anoplocephala perfoliata, the lappeted tapeworm, is an inhabitant of the intestine of equids. Adult tapeworms attach to the intestinal mucosa at the ileocaecal valve and, when present in large numbers, cause edema and hypertrophy of the ileum. The disease manifest by this infection may be inapparent or may give rise to colic (abdominal pain) in the horse apparently from mechanical obstruction or intussusception of the small intestine into the caecocolon.^{1, 2, 3, 4, 5, 6, 7 and 8} The prevalence of infection is geographically variable^{9, 10, 11, 12 and 13} but appears to be increasing,¹⁴ with a much higher rate of infection found with necropsy as opposed to fecal observations. Horses become infected by the ingestion of infected orbatid mites in pastures. Orbatid mites, the intermediate hosts, are predatory and are found in decaying organic material, such as leaf litter. Horses of all ages are infected, but there are lower numbers of clinical cases in horses older than 4 years of age.⁴ The intensity of infection is highest in the late summer and autumn.^{8 and 12} Anthelmintics with reported efficacy against A perfoliata include pyrantel pamoate at 13.2 mg/kg,¹⁰ pyrantel tartrate at 2.6 mg/kg for 30 days,¹⁵ pyrantel embonate at 38 mg/kg,¹⁶ and praziquantel at 1 to 2 mg/kg.^{17 and 18} Nitazoxanide has not been evaluated for Anoplocephala but was included in the trial because of its effects against nematodes and tapeworms in humans.¹⁹ Because Anoplocephala infections may cause disease and there is a perception that current anthelmintics may not be as effective as in the past, a study was done to compare anthelmintics to lower the intensity of fecal egg counts in a herd of horses in central Texas.

Materials and methods

Quarter horse mares and weanlings from a single herd were evaluated with 5 g of feces with a sucrose double centrifugation test to determine whether eggs of Anoplocephala were present.²⁰ Feces from each individual horse were evaluated twice, once approximately 2 weeks before treatment and again on the day of treatment. If Anoplocephala eggs were found on either date, the horse was considered to have positive results. Within each group (mares or weanlings), the treatment selection was randomly allocated as the horses were restrained for treatment. Fecal samples were again evaluated at 14 and 28 days after treatment for the presence or absence of eggs on either day.The dose for each individual horse was determined by chest girth weight tape at the time of treatment. The treatments were as follows: pyrantel pamoate (Strongid-T, Pfizer Animal Health, Exton, Pa) at 13.7 mg/kg via nasogastric intubation (12 mares, 8 weanlings), nitazoxanide oral paste (Nitazoxanide, Idexx Laboratories, Westbrook, Me) at 100 mg/kg (9 mares, 8 weanlings), praziquantel (Droncet injectable, Bayer Corp, Shawnee Mission, Kan) at 1.23 mg/kg via nasogastric intubation (6 mares), and untreated controls (6 mares, 6 weanlings). A 1-tailed Fisher exact test was used to compare rates of infection before and after treatment. If a mare or foal did not have positive results before treatment, it was not evaluated in this study.

Results and discussion

No abnormal clinical signs were seen after treatment with any of the products. Treatment was administered to several additional animals with each product, but they were not included in the analysis if they did not have positive results on 1 of the 2 evaluations before treatment, hence, the different numbers of horses in treatment groups.None of the horses in the trial exhibited clinical signs associated with the infection of A perfoliata. However, before the trial, a mare from the infected herd exhibited signs of colic and Anoplocephala eggs were detected in the feces. Examination of the remainder of the herd gave impetus to the study.Mean egg counts before and after treatment are given in the Table.The presence of strongylate and Parascaris eggs in weanlings served as a control of the methodology of evaluation. The difficulty of finding Anoplocephala eggs has been recognized by several authors,^{5, 8, 13, 14 and 21} but the authors also recognize that when there were greater numbers of parasites there was increased egg production. Therefore, finding of eggs with fecal flotation indicated that there were 20 worms or more. However, there appears to be no correlation between the number of worms and egg counts once the detection threshold is reached,²² so the criterion for evaluation was the presence of eggs in the feces before treatment compared with after treatment. Although mean egg counts were not compared, the number of eggs in each infected horse was less after treatment in all groups compared with untreated controls (Table). The method of evaluation used in this study cannot be equated to those of critical^{10 and 16} or control¹⁴ studies in which horses are killed so that all worms are detected. However, the use of clinical studies to compare compounds is useful in detecting which anthelmintics are likely to be of value against geographically distinct populations of worms. Admittedly, more sampling may have increased the number of horses with positive results, both before and after treatment.     

Hereditary multifocal renal cystadeno-carcinomas and nodular dermatofibrosis in 51 German sheoherd does

The clinical findings In German shepherd dogs with hereditary multifocal renal cystadenocarcinomas and nodular dermatofibrosis are presented. Between 1978 and 1996, 51 cases were examined. Eight cases were detected after being offered a clinical examination because the disease was present in a parent. The remaining 43 dogs were diagnosed after an unsolicited visit to a clinic because of a specific problem. Skin lesions were the main reason (37 per cent) the owners presented their dog for examination. The mean age at diagnosis of renal cystadenocarcinomas and nodular dermatofibrosis was 8-2 years. The male-to-female ratio was 1-1, while the corresponding figure for a reference population was 1–25. Enlarged and abnormally shaped kidneys were palpated In 60 per cent of the dogs and were detected by radiography in 86 per cent of cases. The renal lesions, including metastases, were the main reason for euthanasia and death. The mean age at death was 9-3 years, and the mean age at the first detection of nodular dermatofibrosis was 6-4 years.     

Ryegrass-based diet and barley supplementation: partition of nitrogenous nutrients among splanchnic tissues and hind limb in finishing lambs

Splanchnic metabolism of nitrogenous nutrients and their uptake by the hind limb were studied in finishing lambs receiving ryegrass harvested at grazing stage with or without barley supplementation. Six multicatheterized lambs (40.2 +/- 1.5 kg) were fed with frozen ryegrass (RG) at 690 kJ of ME intake (MEI) x d(-1) x BW(-0.75) and 20.8 g of N intake (NI)/d successively without and with barley supplementation (RG + B), according to a crossover design. Barley supplementation represented 21% of DM intake and increased the MEI and the NI by 32 and 24% respectively, (P < 0.01). In the ruminal fluid, barley increased acetate and butyrate concentrations by 21.2 and 49.6%, respectively (P < 0.04), without any effect on the ammonia concentration. Consequently, the net portal appearance (NPA) of ammonia was not modified, but the NPA of total amino acids (TAA; +38%) and nonessential amino acids (NEAA; +45%) was increased (P < 0.05) by barley supplementation. Taken individually, the NPA of the essential amino acids (EAA) was increased for isoleucine (+32%; P < 0.05), threonine (+151%; P < 0.03), and lysine (+26%; P < 0.06), with no effect for the other EAA. In contrast to what was observed at the PDV level, no significant alteration in the net hepatic amino acid flux was observed for TAA, EAA, NEAA, branched-chain amino acids (BCAA), urea, and ammonia after barley supplementation, showing a relatively minor role of the liver in the regulation of the supply of amino acids to the peripheral tissues. However, taken individually, the net hepatic uptake of some NEAA involved in gluconeogenesis and/or ureagenesis was altered with barley supplementation: the alanine uptake was increased by 44% (P < 0.05), aspartate + asparagine (asx) uptake was decreased by 18% (P < 0.01), and glutamate + glutamine (glx) release tended (P < 0.10) to be increased by 208%. With barley supplementation, NI increased by 5 g of N/d, and net splanchnic release increased by 4.63 g of N/d. Consequently, the additional dietary N supply (together with energy supply) was nearly exclusively available to peripheral tissues as AA-N (N as amino acids), but no strong effect of this additional supply of AA to the hind limb could be demonstrated in terms of net AA hind limb fluxes. Consequently, barley supplementation of a ryegrass-based diet increased the net AA release by the splanchnic tissues, with little effect on the AA net uptake by the peripheral tissues.     

Influence of dietary cobalt source and concentration on performance, vitamin B12 status, and ruminal and plasma metabolites in growing and finishing steers

Sixty Angus steers, averaging 274 kg, were used to evaluate the effects of Co source and concentration on performance, vitamin B12 status, and metabolic characteristics of steers. Treatments consisted of 0 (control, analyzed 0.04 mg Co/kg), 0.05, 0.10, and 1.0 mg of supplemental Co/kg of DM from CoCO3 or 0.05 and 0.10 mg of supplemental Co/kg of DM from Co propionate. Steers were individually fed a growing diet for 56 d followed by a high-concentrate finishing diet. Performance was not affected by Co supplementation during the growing phase. During the finishing phase, ADFI (DM basis) and ADG were higher (P < 0.05) for the entire finishing phase, and gain:feed was higher (P < 0.10) over the first 56 d for Co-supplemented steers. Steers supplemented with 0.10 mg Co/kg as Co propionate had higher (P < 0.05) ruminal propionate and lower (P < 0.05) acetate molar proportions than steers receiving 0.10 Co/kg as CoCO3 during the growing phase. Supplemental Co increased (P < 0.10) molar proportion of propionate during the finishing phase. Plasma vitamin B12 was higher (P < 0.05) in Co-supplemented steers by d 56 of the growing phase and remained higher (P < 0.10) throughout the study. Control steers had higher (P < 0.05) plasma methylmalonic acid on d 56 of the growing phase and on d 28, 56, and 112 of the finishing phase than steers receiving supplemental Co. Steers supplemented with Co had higher plasma glucose at d 56 (P < 0.01), 84 (P < 0.10), and 112 (P < 0.01) of the finishing phase. Steers supplemented with 0.10 mg Co/kg as Co propionate had higher plasma glucose than those receiving 0.10 mg Co/kg as CoCO3 at d 28 of the growing phase (P < 0.05) and d 28 of the finishing phase (P < 0.10). Final body weight and hot carcass weight were lower (P < 0.10) in steers receiving the control diet, whereas other carcass characteristics were not affected by dietary Co. Average daily gain and feed efficiency for the entire finishing phase did not differ among Co-supplemented steers. However, increasing supplemental Co above 0.05 mg/kg DM (total diet Co = 0.09 mg/kg) resulted in increased (P < 0.01) plasma (linear) and liver (quadratic) vitamin B12 concentrations and decreased (quadratic, P < 0.10) plasma methylmalonic acid concentrations toward the end of the finishing phase. These results suggest that finishing steers require approximately 0.15 mg Co/kg of DM. Vitamin B12 status was not affected by Co source; however, the two Co sources seemed to affect certain metabolites differently.