Severe biliary hyperplasia associated with liver fluke infection in an adult alpaca.

An adult alpaca (Lama pacos) had a locally extensive area of hepatic atrophy involving the right lobe. Grossly, the atrophic lobe was light tan and firm and contained small, raised, white to yellow, partially mineralized circular nodules predominantly at the periphery of the atrophic tissue. Microscopically, viable hepatocytes were not present in the atrophic area, and the tissue consisted of diffuse biliary epithelial proliferation without any evidence of nuclear or cellular atypia or the presence of mitotic figures. The circular mineralized nodules consisted of granulomatous inflammation with intralesional parasitic ova surrounded by fibrous connective tissue. Morphologically, the ova were compatible with those of Fasciola hepatica. The severe biliary hyperplasia was unusual, and it was not clear whether it was caused by an aberrant host response to the parasitic infection or whether it was an unrelated event.

Seasonal changes in serum calcium, phosphorus, and vitamin D concentrations in llamas and alpacas.

OBJECTIVE: To evaluate the interaction of season and age on serum calcium, phosphorus, and vitamin D3 concentrations in llamas and alpacas. ANIMALS: 23 clinically normal llamas and 7 alpacas. PROCEDURES: Animals were assigned to 1 of the 3 following groups on the basis of age at the start of the study: adult (age, > or = 24 months; n = 8), yearling (> 12 but < 20 months; 5), and neonate (< 6 months; 17). Twelve serum samples were obtained at monthly intervals. Calcium, phosphorus, and vitamin D3 concentrations were measured, and the calcium-to-phosphorus concentration (Ca:P) ratio calculated. Effect of season and age on each of these variables was determined. RESULTS: Vitamin D3 concentrations varied significantly as a function of season; the highest and lowest concentrations were detected September through October and February through March, respectively. The seasonal decrease in vitamin D3 concentration was significantly greater in neonates and yearlings, compared with adults. Serum phosphorus concentration decreased as a function of age, with the most significant seasonal change detected in the neonate group. The Ca:P ratio in neonates varied between 1.1 and 1.3 except during winter months when it increased to > or = 2.0. CONCLUSIONS AND CLINICAL RELEVANCE: Mean vitamin D3 concentration varied by > 6 fold in neonatal and yearling llamas and alpacas and > 3 fold in adult animals as a function of season. These results support the hypothesis that seasonal alterations in vitamin D3 concentrations are a key factor in the development of hypophosphatemic rickets in llamas and alpacas.

Metabolic changes and induction of hepatic lipidosis during feed restriction in llamas.

OBJECTIVES: To determine whether feed restriction induces hepatic lipidosis (HL) in llamas and to evaluate the metabolic changes that develop during feed restriction. ANIMALS: 8 healthy adult female llamas. PROCEDURE: Llamas were fed grass hay at a rate of 0.25% of their body weight per day for 13 to 28 days. Llamas were monitored by use of clinical observation, serum biochemical analyses, and ultrasound-guided liver biopsies. RESULTS: All 8 llamas lost weight and mobilized fat. Five llamas developed HL, including 4 that were nursing crias. During the period of feed restriction, mean serum concentration of bile acids and activities of aspartate aminotransferase (AST), sorbitol dehydrogenase (SDH), and gamma-glutamyl transferase (GGT) were significantly higher in llamas that developed HL, compared with llamas that did not. Mean insulin-to-cortisol concentration ratios were lower in llamas with HL before and up to 7 days of feed restriction, compared with those that did not develop HL. CONCLUSIONS AND CLINICAL RELEVANCE: HL in llamas may be induced by severe feed restriction, particularly in the face of increased energy demand. Llamas with weight loss attributable to inadequate dietary intake may develop biochemical evidence of hepatopathy and HL. Increases in serum concentration of bile acids and activities of GGT, AST, and SDH may indicate the development of HL in llamas and identify affected animals for aggressive therapeutic intervention.

Glucose tolerance testing in llamas and alpacas.

OBJECTIVE: To determine blood glucose clearance in 2 species of New World camelids after IV challenge and to examine mechanisms of this clearance. ANIMALS: 5 adult female llamas and 5 adult gelded alpacas. PROCEDURE: After food was withheld for 12 hours, camelids received 0.5 g of glucose/kg of body weight by rapid IV infusion. Serum concentrations of glucose, nonesterified fatty acids, cortisol, and insulin, and plasma concentrations of lactate were determined before and 0, 1, 2, 3, 4, 5, 15, 30, 60, 90, 120, 180, and 240 minutes after infusion. Ratios of insulin to glucose and insulin to cortisol were calculated for each time point. RESULTS: Postinfusion glucose concentrations were significantly higher in llamas than alpacas for the first 15 minutes and remained significantly higher than baseline values in both species for 180 minutes. Lactate and cortisol concentrations did not change significantly; nonesterified fatty acid concentrations decreased in both species 30 minutes after infusion. Baseline insulin concentrations were < 6 microU/ml in both species and increased only to 10.1 +/- 0.7 microU/ml in llamas. Insulin concentrations did not change significantly in alpacas. CONCLUSIONS AND CLINICAL RELEVANCE: Llamas and alpacas clear glucose more slowly than other domestic species after challenge, mainly because of a weak insulin response and slow cellular uptake. This response may impair the assimilation of exogenous glucose as well as make llamas and alpacas prone to diabetes-like disorders when an abundance of endogenous or exogenous glucogenic agents are present.

Pharmacokinetics and pharmacodynamics of antiulcer agents in llama.

Plasma concentration time curves following intravenous (i.v.) administration of 1.5 mg/kg of ranitidine, 0.2 mg/kg, 0.4 mg/kg and 0.8 mg/kg of omeprazole, respectively, were analysed in six llamas. Plasma profiles after i.v. administration of both drugs showed plasma concentrations declining in a biexponential manner with a rapid distribution phase. Pharmacokinetics parameters after ranitidine administration to six llamas showed a mean elimination half-life of 1.53 +/- 0.26 h. The mean volume of distribution (Vdss) in llamas was 1.77 +/- 0.31 L/kg, and mean body clearance in llamas was 0.778 +/- 0.109 L/kg/h. Ranitidine produced only a small transitory (<1 h) decline in acid production when administered i.v. at a dose of 1.5 mg/kg. Omeprazole showed dose-dependent nonlinear pharmacokinetics. The mean half-life of 0.2 mg/kg i.v. omeprazole was shorter than that of 0.4 and 0.8 mg/kg i.v. omeprazole, i.e. 0.61, 0.72 and 1.07 h, respectively. The area under the curve (AUC) and mean residence time (MRT) increased with increasing dose, while clearance decreased as dose increased. The decline in acid production following 0.2 mg/kg i.v. omeprazole was highly variable and did not produce a clinically useful suppression of third compartment acid production. In contrast, both 0.4 mg/kg and 0.8 mg/kg omeprazole i.v. administration significantly reduced third compartment acid production. The reduction in acid production following 0.8 mg/kg omeprazole was not significantly greater than the reduction observed following 0.4 mg/kg dosage. Misoprostol (10 microg/kg) was administered i.v. in an absolute alcohol solution. Two animals collapsed following drug administration. While the side-effects could have been produced by either misoprostol or the alcohol vehicle, the clinical changes were more consistent with an adverse drug reaction. Unfortunately, the limitation of UV detection did not provide the sensitivity needed to quantify the amount of misoprostol in llama plasma, and the pharmacokinetics could not be evaluated.

Pathogenesis of Streptococcus zooepidemicus infection after intratracheal inoculation in llamas.

OBJECTIVES: To test whether generalized Streptococcus zooepidemicus infection could be induced by intratracheal inoculation in llamas and to characterize this infection. ANIMALS: 6 test and 3 control llamas. PROCEDURE: Test llamas received 1 of 3 dosages of S. zooepidemicus by intratracheal injection, whereas control llamas received sterile culture medium. Physical examination variables and results of clinicopathologic analyses of blood, peritoneal fluid, and tracheal wash fluid were compared in test llamas between, before, and during the development of bacteremia and with control llamas. Bacteriologic culture was performed on all collected body fluids and tissue specimens that were collected at necropsy. Tissue specimens that were collected at necropsy were examined histologically. RESULTS: Infection induced fever, anorexia, and signs of depression. Five of 6 infected llamas developed specific signs of inflammation in the thorax or abdomen, bacteremia, neutrophilic leukocytosis with toxic changes and high band neutrophil cell counts, hyperfibrinogenemia, and high peritoneal fluid WBC counts and protein concentrations. On development of bacteremia, llamas had significant decreases in serum iron (from 118+/-25 to 6+/-4 microg/ml) and increases in serum glucose (from 131+/-5 to 253+/-48 mg/dl) concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: Streptococcus zooepidemicus spreads rapidly to other body compartments after intratracheal inoculation in llamas. Fever, anorexia, and signs of depression are the most consistent clinical signs, although other signs are possible. Clinicopathologic analysis of body fluids yields evidence of inflammation. Infection by S. zooepidemicus can be proven by bacteriologic culture of body fluids before death or of tissue specimens after death.

Use of cloprostenol as an abortifacient in the llama (Lama glama).

As part of a larger project investigating the development and heritability of choanal atresia glama), it was necessary to develop a protocol for aborting llamas at various stages of gestation. Twenty-seven animals between 4 and 7 mo of gestation were successfully aborted a total of 53 times following two 250 microg intramuscular injections of cloprostenol at 24 h intervals. Abortion was induced once in 10 animals and multiple times (range 2 to 5) in 17 animals. Twenty-four animals (45.2%) aborted 3 d following the first injection, with 20 animals (37.7%) aborting 4 d post prostaglandin administration. Other animals aborted at 2 d (n=6, 11.3%), 5 d (n=2, 3.8%), and 7 d (n=1, 1.9%) following drug administration. Forty-nine (92.5%) of the abortions occurred following a single series of injections, while 4 animals (7.5%) aborted following a second series of injections. No confirmed pregnant animals failed to abort following the second series of cloprostenol injections. Conception rates in animals rebred 2 to 4 wk following an abortion were comparable to those of untreated animals in the research herd. Unlike the severe hypertension and death that has been reported following dinoprost tromethamine administration in the llama, no adverse reactions were observed in this study following cloprostenol administration. The results demonstrate that llamas can be safely and effectively aborted up to 7 mo of gestation (normal full term gestation = 342 +/- 10 days) without adverse effects on subsequent fertility.

Specificity of four serologic assays for Mycobacterium avium ss paratuberculosis in llamas and alpacas: a single herd study.

An investigation was conducted for Mycobacterium avium ss paratuberculosis infections in a research herd of llamas and alpacas. Herd culture-negative status was established over a 23-month period by screening any individuals with any signs compatible with paratuberculosis (n = 1), high serology values (n = 8), or other health and research related reasons (n = 24). There were no M. avium ss paratuberculosis isolates from radiometric cultures of multiple tissue and fecal samples from these individuals and no known sources of exposure. Paratuberculosis is uncommon in North American llamas and alpacas: only 5 cases were identified after an extensive search of the Veterinary Medical Data Base, diagnostic laboratory records, publication databases, and personal communications. Therefore, serum samples from llamas (n = 84) and alpacas (n = 16) in the culture-negative herd were used to obtain preliminary estimates of test specificity for 3 enzyme-linked immunoassays (ELISAs) and an agar gel immunodiffusion (AGID) assay kit for detecting serum antibodies to M. avium ss paratuberculosis in South American camelids. The ELISAs were modifications of established bovine assays for antibody detection. With provisional cutoffs, ELISA-A had 52 false positives (specificity 48%), ELISA-B had 8 false positives (specificity 92%), ELISA-C had two false positives (specificity 98%), and the AGID had 0 false positives (specificity 100%). The range of ELISA values for culture-positive llamas and alpacas (n = 10) from other herds overlapped the range of values for culture-negative llamas and alpacas. The accuracy of the ELISAs may be improved by using age- and sex-specific cutoffs because uninfected male llamas and alpacas that were older than 1 year had higher values for some tests. These tests can be used for either llamas or alpacas; the protein-G conjugate ELISA (ELISA-B) may be useful for multispecies applications. These assays are best used for rapid presumptive diagnoses of llamas and alpacas with diarrhea and weight loss and as a screening tool for herds known to be exposed to infection. All seropositive results should be confirmed with culture.

Radiographic contrast gastrointestinal study of the neonatal llama.

This is a description of the radiographic appearance of the normal gastrointestinal tract of neonatal crias with survey and contrast radiography, including transit times for the various segments of the gastrointestinal tract. Radiographs of the abdomen of six healthy neonatal llama crias positioned in right lateral and dorsal recombency were obtained initially then at various intervals post barium administration. Portions of the gastrointestinal tract that could be identified included the stomach with first, second and third compartments, the duodenal ampulla, small intestine, proximal loop of the ascending colon, spiral colon and descending colon. Barium given by orogastric intubation rapidly entered all three compartments of the stomach and the proximal small intestine. Overall the transit time through the intestinal tract of the crias was longer than expected with no emptying of the ascending colon and beyond though studies were carried to 48-72 hours. Recommendations are made for frequency of radiographs.

Hepatic lipidosis in llamas and alpacas: 31 cases (1991-1997).

OBJECTIVE: To identify factors associated with hepatic lipidosis (HL) in llamas and alpacas. DESIGN: Retrospective case series. ANIMALS: 30 llamas and 1 alpaca. PROCEDURES: Medical records were searched to identify llamas or alpacas in which a histologic diagnosis of HL was made. Information was retrieved on signalment, history, clinical and laboratory findings, and results of necropsy or examination of biopsy specimens. Data were analyzed using descriptive statistics and chi 2 analyses. RESULTS: Females were affected more often than males; however, the sex distribution was not different from that of the camelid population in the diagnostic laboratory's database. Fifty-four percent of the females were pregnant, and 46% were lactating. Most affected camelids were 6 to 10 years old. Anorexia and recent weight loss were common (51.6% of camelids). An infective agent was found in only one ilama, and toxins and mineral deficiencies were not identified. The most common abnormalities on serum biochemical analysis were a high concentration of bile acids, high activities of gamma-glutamyltransferase (GGT) and aspartate aminotransferase (AST), and hypoproteinemia. Concentrations of nonesterified fatty acids (NEFA) and beta-hydroxybutyrate (beta-HB) were high in those camelids in which these compounds were assayed. Twenty-nine camelids did not survive. CLINICAL IMPLICATIONS: Sick camelids should be considered at risk for developing HL, especially those with anorexia or the metabolic demands of pregnancy and lactation. Other stresses also appear to contribute. High concentrations of NEFA, beta-HB, and bile acids; high activities of GGT and AST; and hypoproteinemia may indicate that HL has developed.

Blood mineral and vitamin E concentrations in llamas.

OBJECTIVE: To establish reference values for blood concentrations of total calcium (Ca), inorganic phosphorus (P), iron (Fe), copper (Cu), zinc (Zn), selenium (Se), and vitamin E (Vit E) in clinically normal llamas. ANIMALS: 270 llamas ranging in age from < 1 month to > 15 years and grouped by age, sex, pregnancy status, and stage of gestation. Selected llamas were from 21 farms in Oregon, did not have previous health problems, and met specific health criteria on examination. PROCEDURE: Serum and blood samples were obtained and analyzed for concentrations of Ca, P, Fe, Cu, Se, Zn, and Vit E, and total iron binding capacity (TIBC) and percentage of transferrin saturation (% Sat). Mean differences by age, sex, pregnancy status, and stage of gestation, as well as all interactions, were compared to establish reference values. RESULTS: Mean values and reference ranges for most of the minerals and vitamins were similar to previously reported values. Male versus female differences were not identified for any measurements. Age was a significant variable for Ca, P, Fe, and Se concentrations, as well as Ca-to-P ratio and TIBC. Identified age-based effects were modeled by use of linear regression. Copper and Zn concentrations and % Sat did not differ as a function of age. Serum Vit E concentration was influenced by an age by sex interaction and stage of gestation. CONCLUSIONS: Age was found to be an important variable influencing many blood nutrient concentrations in healthy llamas. CLINICAL RELEVANCE: Clinical diagnosis of metabolic disease may be improved with use of age-based reference values, especially for neonates.

An overview of selected diseases and drug needs in the llama and alpaca industries.

The North American llama and alpaca industries will probably continue to expand at a rate of 20-25% per year for at least the next one to two decades. As a result, while the total population will always be small relative to the cattle and swine industries, the total numbers of alpacas and llamas will be substantial. Accordingly, it is appropriate and timely to begin the process of getting drugs approved for usage in these species. Four drugs that warrant consideration for NRSP-7 evaluation and approval are: Ivermectin for the prevention of P. tenuis infections. Chlorsulon for the treatment of Fasciola hepatica infections. Ceftiofur sodium for the treatment of various bacterial infections. Omeprazole for the management and prevention of third compartment ulcers.

Normal reference intervals and the effects of time and feeding on serum bile acid concentrations in llamas.

Fifty clinically healthy llamas, 0.5-13 years of age (22 intact males, 10 neutered males, 18 females), with no biochemical evidence of liver disease or hematologic abnormalities, were selected to establish serum bile acid reference intervals. Serum samples submitted to the clinical pathology laboratory were analyzed using a colorimetric enzymatic assay to establish bile acid reference intervals. A nonparametric distribution of llama bile acid concentrations was 1-23 micromol/liter for llamas >1 year of age and 10-44 micromol/liter for llamas < or = 1 year of age. A significant difference was found between these 2 age groups. No correlation was detected between gender and bile acid concentrations. The reference intervals were 1.1-22.9 micromol/liter for llamas >1 year of age and 1.8-49.8 micromol/liter for llamas < or = 1 year of age. Additionally, a separate group of 10 healthy adult llamas (5 males, 5 females, 5-11 years of age) without biochemical or hematologic abnormalities was selected to assess the effects of feeding and time intervals on serum bile acid concentrations. These 10 llamas were provided fresh water and hay ad libitum, and serum samples were obtained via an indwelling jugular catheter hourly for 11 hours. Llamas were then kept from food overnight (12 hours), and subsequent samples were taken prior to feeding (fasting baseline time, 23 hours after trial initiation) and postprandially at 0.5, 1, 2, 4, and 8 hours. In feeding trials, there was no consistent interaction between bile acid concentrations and time, feeding, or 12-hour fasting. Prior feeding or time of day did not result in serum bile acid concentrations outside the reference interval, but concentrations from individual llamas varied within this interval over time.

The disposition of five therapeutically important antimicrobial agents in llamas.

The disposition of five therapeutic antimicrobial agents was studied in llamas (Lama glama) following intravenous bolus administration. Six llamas were each given ampicillin, tobramycin, trimethoprim, sulfamethoxazole, enrofloxacin and ceftiofur at a dose of 12 mg/kg, 1 mg/kg, 3 mg/kg, 15 mg/kg, 5 mg/kg, and 2.2 mg/kg of body weight, respectively, with a wash out period of at least 3 days between treatments. Plasma concentrations of these antimicrobial agents over 12 h following i.v. bolus dosing were determined by reverse phase HPLC. Disposition of the five antimicrobial agents was described by a two compartment open model with elimination from the central compartment, and also by non-compartmental methods. From compartmental analysis, the elimination rate constant, half-life, and apparent volume of distribution in the central compartment were determined. Statistical moment theory was used to determine noncompartmental pharmacokinetic parameters of mean residence time, clearance, and volume of distribution at steady state. Based on the disposition parameters determined, and stated assumptions of likely effective minimum inhibitory concentrations (MIC) a dose and dosing interval for each of five antimicrobial agents were suggested as 6 mg/kg every 12 h for ampicillin; 4 mg/kg once a day or 0.75 mg/kg every 8 h for tobramycin; 3.0 mg/kg/15 mg/kg every 12 h for trimethoprim/sulfamethoxazole; 5 mg/kg every 12 h for enrofloxacin; and 2.2 mg/kg every 12 h for ceftiofur sodium for llamas. Steady-state peak and trough plasma concentrations were also predicted for the drugs in this study for llamas.

Evaluation of vitamin D status of llamas and alpacas with hypophosphatemic rickets.

OBJECTIVE: To evaluate vitamin D concentrations in juvenile llamas and alpacas with hypophosphatemic rickets. DESIGN: Prospective cohort study. ANIMALS: 21 llamas (14 with rickets, 7 clinically normal) and 9 alpacas (6 with rickets, 3 clinically normal). PROCEDURES: Blood samples were collected at the time of diagnosis and prior to the initiation of treatment. Serum concentrations of calcium, inorganic phosphorus, and 25-hydroxycholecalciferol (vitamin D3) were determined on all samples. Comparisons were completed for disease status, age, sex, species, month of birth, and all interactions. RESULTS: Serum concentrations of phosphorus and vitamin D were lower in affected llamas and alpacas than in clinically normal llamas and alpacas, even when mean concentrations were adjusted for age differences. Species (llama or alpaca), sex, and age did not affect any of the metabolite concentrations within this study population. Month of birth influenced vitamin D concentrations and number of affected llamas and alpacas per month. The greatest number of affected llamas and alpacas was identified between January through March, suggesting a seasonal pattern to this syndrome. Treatment of affected llamas and alpacas with vitamin D resulted in increased concentrations of phosphorus and vitamin D. Serum phosphorus concentration was best predicted by 2 independent variables (serum vitamin D concentration and month of birth). CLINICAL IMPLICATIONS: We believe vitamin D deficiency is the primary cause of hypophosphatemic-rickets of growing camelids, and the observed hypophosphatemia is secondary to a primary deficiency of vitamin D. Appropriate treatment with vitamin D supplements can correct hypophosphatemia and vitamin D deficiency in camelids.

Megaesophagus in 15 llamas: a retrospective study (1985-1993).

Megaesophagus was diagnosed in 15 llamas on the basis of survey and contrast radiography. Age of onset ranged from 13 months to 9.5 years. Clinical signs varied, with salivation being the most common; regurgitation or dysphagia were noted in only 33% of the cases. Duration of the disorder ranged from 1 week to 5 years. Organophosphate toxicity was the cause of megaesophagus in 1 llama. While most cases were of unknown etiology, the 3 with histological abnormalities included 1 with vagal neuropathy and 2 with a degenerative myopathy of esophageal muscles. The causes in most were undetermined.

Third compartment ulcers in the llama.

The diagnosis of third compartment ulcers in the llama and alpaca is largely one of exclusion. Clinical signs may include mild to severe colic, inappetence, decreased fecal output, bruxism, and depression. Abdominocentesis results are usually unremarkable if C3 perforation has not occurred but reflective of a generalized peritonitis if full thickness ulceration has occurred. The H-2 receptor antagonists cimetidine and ranitidine do not suppress C3 acid production for a significant period of time and are of questionable efficacy in the management of C3 ulcers.

Suppression of prolactin and cytotoxic T-lymphocyte activity in PCB-treated mice.

Halogenated aromatic hydrocarbons (HAH) are ubiquitous environmental contaminants. Studies in rats have shown that HAH treatment can lead to dysregulation of circulating hormone levels, including prolactin. Reduction of prolactin levels in both rats and mice is inhibitory to immune function. Previous studies have reported suppression of alloantigen-specific cytotoxic T-lymphocyte (CTL) activity in mice treated with 3,3', 4,4', 5,5'-hexachlorobiphenyl (HxCB). Here we report that treatment of mice with HxCB (10 mg/kg body weight) leads to a significant reduction of serum prolactin levels (by 89% to 3.7 ng/ml) on day 10 post alloantigen injection (P815 mastocytoma), the day of peak alloantigen-specific CTL activity. Prolactin levels were not altered on day 3 post alloantigen injection. Treatment with bromocriptine (5 mg/kg/day) reduced serum prolactin levels slightly on day 3 and significantly (94% to 2.1 ng/ml) on day 10 post alloantigen injection. Splenic CTL activity was not altered by treatment with bromocriptine. The data presented here suggest that reduction of prolactin levels alone, to the extent observed in HxCB-treated mice, is not causative for CTL suppression.