Different isolates from Leishmania braziliensis complex induce distinct histopathological features in a murine model of infection

The aim of this study was to evaluate the histopathological features in tissues of mice infected by human isolates (I, II, and III) or the reference M2903 strain of Leishmania braziliensis complex. BALB/c and C57Bl/6 mice were infected in the hind footpad with 10⁶ stationary-phase promastigotes of L. braziliensis complex. The evolution of lesions was observed for 10 weeks and the animals were then euthanized and liver, spleen and popliteal lymph nodes were collected. Tissues were stained with hematoxylin and eosin and analyzed by immunohistochemistry assay. Increased thickness of infected footpads was observed in all animals, lesions were nodular and non-ulcerated. Mice infected with isolate I presented inflammatory infiltrates consisting predominantly of mononuclear cells in all tissues examined, and also a great number of megakaryocytes, compared with other isolates. Infection with isolate II led to an infected footpad enlargement not seen in other isolates. In addition, mononuclear infiltrates in the liver and hemosiderin in spleen were noted. Conversely, mice infected with either isolate III or M2903 strain only showed an increased number of megakaryocytes in spleen. All tissues examined had detectable amastigote forms of Leishmania by immunohistochemistry in all groups. Taking together, our results showed an unforeseen behavior of different isolates of L. braziliensis complex that led to diverse pathological findings.     

Cytomorphologic differentiation of benign and malignant mammary tumors in fine needle aspirate specimens from irradiated female Sprague-Dawley rats.

BACKGROUND: Fine needle aspiration (FNA) offers a rapid and minimally invasive means to distinguish malignant from benign neoplasms. However, few studies have been published regarding the cytopathology of mammary tumors in rats despite widespread use of the rat model for breast cancer formation and inhibition. OBJECTIVE: The purpose of this study was to determine the diagnostic accuracy of FNA cytology and to develop distinguishing cytologic criteria for the diagnosis of radiation-induced benign and malignant mammary tumors in rats. METHODS: In a study of radiation-induced mammary carcinogenesis, 100 Sprague-Dawley rats with cutaneous masses were randomly chosen for FNA. The aspirates were smeared, fixed, and stained with a modified Papanicolaou procedure for diagnostic evaluation. Cytologic and histologic diagnoses (benign vs malignant) were compared, and diagnostic accuracy was calculated using the histologic diagnosis as the criterion standard. FNA smears were scored semiquantitatively on a scale of 1-4 for cellularity, atypia, nuclear size, chromatin pattern, nuclear membrane thickness, nucleoli, and mitoses. The background was evaluated for necrosis, hemorrhage, inflammation, and mucosecretory material. Cytomorphologic features were compared statistically between benign and malignant tumors, based on the histologic diagnosis. RESULTS: The sensitivity of FNA was 92.3% and specificity was 89.4% for the detection of malignancy. However, 14% of specimens, all fibroadenomas by histology, had insufficient cells for cytologic evaluation, for an overall accuracy rate of 78.0%. Malignant tumors had significantly higher scores for all cytomorphologic features, and were significantly more likely to contain cell clusters and necrotic debris. CONCLUSIONS: FNA is an accurate method for differentiating benign and malignant rat mammary tumors.     

Effects of cereals and/or protein supplement extrusion on diet utilisation and performance of intensively reared cattle

The effect of extruding the cereal and/or the protein supplement of a compound feed on its nutritive value and on the performance of intensively reared male calves was studied. The compound feed was formulated with 0.65 of a cereal blend (60:40 maize:barley), 0.25 of a protein blend (1/3:1/3:1/3 raw soybeans:peas:lupins), and 0.08 of urea to contain 0.17 of crude protein. It was tested without extruding (NE) and with the cereal blend (CE), the protein blend (PE) or both (CPE) extruded. Another non-extruded compound feed with mainly soybean meal as the protein supplement (NE-SBM) also was studied. The five experimental compound feeds together with barley straw were offered ad libitum to 50 male Friesian calves (112 kg initial live weight) for 13 weeks, recording individual feed intake and live weight gain. Crude protein (CP) solubility and in vitro and in situ CP degradation of experimental compounds, and in vitro gas production kinetics, in vivo digestibility and urinary allantoin excretion of diets were determined. Cereal extrusion promoted a greater gas production at all incubation times (p < 0.001), associated to the increase in starch gelatinisation. PE and NE-SBM compounds showed lower in vitro (p < 0.01) and in situ (p < 0.001) CP degradability than the other feeds. Ingredient extrusion did not affect apparent DM, OM and CP digestibilities, but diet CPE showed a lower NDF digestibility (p < 0.05) than the others (0.293 vs. 0.420, 0.387, 0.390 and 0.407 with CPE vs. NE, CE, PE and NE-SBM, respectively). The apparent EE digestibility increased (p = 0.053) with the inclusion of extruded ingredients, (0.799, 0.749 and 0.794 vs. 0.719 and 0.702 with CE, PE and CPE vs. NE and NE-SBM, respectively). Daily allantoin excretion was lower (p < 0.01) in calves receiving the CPE diet than in those receiving the NE, PE and NE-SBM diets. Treatments did not affect the average daily gains (1.61 ± 0.022 kg d^− 1), however calves on the CPE diet showed a lower (p < 0.01) concentrate and total conversion ratios than those fed NE, PE and NE-SBM (2.60 vs. 2.79, 2.85 and 2.98 kg concentrate DM kg^− 1 daily gain and 2.93 vs. 3.09, 3.13 and 3.26 kg total DM kg^− 1 daily gain, respectively). The CE diet resulted in numerically higher (p > 0.05) concentrate and total DM conversion ratios (2.72 and 3.02) than CPE and lower (p < 0.05) than NE-SBM. Improvement in feed conversion ratio after extrusion would be related to a better starch utilisation and not to changes in microbial or dietary protein flow to the duodenum, although a better utilisation of fat energy cannot be disregarded.     

An estimate of the methionine requirement and its variability in growing pigs using the indicator amino acid oxidation technique

Although AA requirements for the mean of a population of growing pigs have been established using traditional methods, there are no estimates of the variability within the population and whether this variation differs among AA. With the increased use of supplemental Lys in pig diets, there will be an increased need to supplement Met, commonly the second or third limiting AA in corn-soybean diets. The indicator AA oxidation method allows repeated measurements in a short period of time so that the AA requirement can be determined for individual pigs at a similar physiological stage. The objective of this study was to determine the mean Met requirement in individual gilts and to estimate the related variability. Six individually housed female pigs (initial BW = 8.8 kg, SD 1.5) each received diets providing 6 levels of dl-Met. The isonitrogenous and isoenergetic diets contained 0.187, 0.250, 0.290, 0.320, 0.350, and 0.377% Met (analyzed, as-fed basis). Cysteine (0.48%) and Lys (1.44%) concentrations were similar for all diets. Pigs were adapted for 6 d to the basal corn-soybean meal diet (0.187% Met), which was offered at 95 g/kg(0.75) of BW to ensure complete consumption of the test diets. During 4-h oxidation studies, 313.4 kBq, (SD 35.6) of L-[1-(14)C]Phe was mixed with each of 8 half-hourly meals, and expired CO(2) was collected. The breakpoint in Phe oxidation, representing the Met requirement, and its variability, was determined using 2-phase linear regression. Phenylalanine oxidation decreased as the Met content increased from 0.187 to 0.29%. Phenylalanine oxidation was not different (P > 0.2) for diets ranging from 0.320 to 0.377% Met. The dietary Met requirement varied from 0.320 to 0.373% for individual pigs. The mean Met requirement for individual pigs was determined to be 0.340% of diet (SD = 0.024%, CV= 7.1%), with 0.340, 0.364, and 0.388% covering the requirement of 50, 66, and 95% of the population, respectively. The present mean population estimate was similar to the recommended dietary Met concentration of 0.325% for pigs of this BW and feed intake. To maximize profitability, Met levels in starter pig diets should be determined, depending on the cost of crystalline Met and the fraction of the population whose requirement is to be met.     

Estimation of genetic parameters for mature weight in Angus cattle

The aim of this study was to estimate genetic parameters for BW of Angus cattle up to 5 yr of age and to discuss options for including mature weight (MW) in their genetic evaluation. Data were obtained from the American Angus Association. Only records from herds with at least 500 animals and with >10% of animals with BW at ≥ 2 yr of age were considered. Traits were weaning weight (WW, n = 81,525), yearling weight (YW, n = 62,721), and BW measured from 2 to 5 yr of age (MW2, n = 15,927; MW3, n = 12,404; MW4, n = 9,805; MW5, n = 7,546). Genetic parameters were estimated using an AIREML algorithm with a multiple-trait animal model. Fixed effects were contemporary group and departure of the actual age from standard age (205, 365, 730, 1,095, 1,460, and 1,825 d of age for WW, YW, MW2, MW3, MW4, and MW5, respectively). Random effects were animal direct additive genetic, maternal additive genetic, maternal permanent environment, and residual. Estimates of direct genetic variances (kg(2)) were 298 ± 71.8, 563 ± 15.1, 925 ± 52.1, 1,221 ± 65.8, 1,406 ± 80.4, and 1,402 ± 66.9; maternal genetic variances were 167 ± 4.8, 153 ± 6.1, 123 ± 9.1, 136 ± 12.25, 167 ± 18.0, and 110 ± 14.0; maternal permanent environment variances were 124 ± 2.9, 120 ± 4.3, 61 ± 7.5, 69 ± 11.9, 103 ± 15.9, and 134 ± 35.2; and residual variances were 258 ± 3.8, 608 ± 8.6, 829 ± 34.2, 1,016 ± 38.8, 1,017 ± 52.1, and 1,202 ± 63.22 for WW, YW, MW2, MW3, MW4, and MW5, respectively. The direct genetic correlation between WW and YW was 0.84 ± 0.14 and between WW and MW ranged from 0.66 ± 0.06 (WW and MW4) to 0.72 ± 0.11 (WW and MW2). Direct genetic correlations ranged from 0.77 ± 0.08 (YW and MW5) to 0.85 ± 0.07 (YW and MW2) between YW and MW, and they were ≥ 0.95 among MW2, MW3, MW4, and MW5. Maternal genetic correlations between WW and YW and MW ranged from 0.52 ± 0.05 (WW and MW4) to 0.95 ± 0.07 (WW and YW), and among MW they ranged from 0.54 ± 0.14 (MW4 and MW5) to 0.94 ± 0.07 (MW2 and MW3). Genetic correlations suggest that a genetic evaluation for MW may be MW2-based and that including BW from older ages could be accomplished by adjusting records to the scale of MW2.     

Studies on the effects of essential-oil-based feed additives on performance, ileal nutrient digestibility, and selected bacterial groups in the gastrointestinal tract of piglets

The aim of this study was to assess the effects of 2 different phytogenic products on performance, ileal nutrient digestibility, and composition of the intestinal microbiota. The 2 phytogenic products contained different essential oil mixtures (EOM) characterized by either menthol (Mentha arvensis; EOM-M) or cinnamon aldehyde (Cinnamomum aromaticum; EOM-C) as main constituents. Three treatments consisted of control diet without EOM addition and diets supplemented with EOM-M or EOM-C. Reproducibility of the effects was examined in 4 trials with a total of 300 male castrated and female piglets weaned at 25 d of age. The number of pens per treatment in trials I through III were 7, 9, and 9, respectively, for research station conditions, and 10 in trial IV for simulated farm conditions. In research station conditions, the experimental unit consisted of flat deck pens with 2 piglets per pen, whereas it consisted of floor pens with straw bedding with 5 piglets per pen in farm conditions. The feed additives had no effect on feed intake or BW gain. Improvements (P < 0.05) in G:F were observed for EOM-M supplemented diets in 2 of 4 trials as well as for the combined data of all trials. These improvements were associated with greater (P<0.05) apparent ileal digestibility of CP and of most AA. The effect of EOM-C on these response criteria was intermediate between control and EOM-M. Real-time PCR analysis of the gastrointestinal contents for 7 bacterial groups (Lactobacillus spp., Enterococcus spp., Clostridium coccoides and Clostridium leptum cluster, Escherichia spp., and Escherichia coli toxin estII) indicated no effect of treatments on the gastrointestinal microbiota. It was concluded that EOM-M consistently improved feed efficiency in weaned piglets, and it was associated with improved ileal protein and AA digestibility. In general, however, the effectiveness of EOM as feed additives differs considerably depending on the constituents.     

Pathologic and immunohistochemical findings in an outbreak of systemic toxoplasmosis in a mob of red kangaroos

Toxoplasma gondii is a zoonotic protozoan pathogen that infects many endothermic vertebrates, including humans; the domestic cat and other felids serve as the definitive host. Macropodids are considered highly susceptible to toxoplasmosis. Here, we describe the clinical, pathologic, and immunohistochemical findings of an outbreak of systemic toxoplasmosis in a mob of 11 red kangaroos (Macropus rufus), with high morbidity (73%) and mortality (100%) rates. Affected animals had either severe and rapidly deteriorating clinical conditions or sudden death, which was correlated with widespread necrotizing lesions in multiple organs and intralesional T. gondii organisms identified via MIC3-specific immunohistochemistry and confirmed by REP529-specific rtPCR. Quantification of parasites demonstrated the highest parasite density in pulmonary parenchyma compared with other tissues. Our study highlights the continued importance of this severe condition in Australian marsupials.     

Osteomalacia as a result of phosphorus deficiency in beef cattle grazing subtropical native pastures in Uruguay

We investigated 2 outbreaks of osteomalacia as a result of phosphorus (P) deficiency in herds of lactating beef cows grazing subtropical native pastures in Uruguay. Cows exhibited pica, difficulty to stand and walk, rib fractures, and body weight loss even with adequate forage availability. Osteopenia and severe osteomalacia were observed on gross and histologic examination. The concentrations of bicarbonate-extractable P in soil (4.0, 4.1 mg P/kg), total P in pasture (0.9, 1.1 g P/kg), inorganic P in serum (1.0, 0.71 mmol P/L), and P in bone (73 mg P/mL) were all low. Although injectable and mineral salt supplements provided additional P in both outbreaks, these supplementary amounts were insufficient to prevent P deficiency. The P ingested by the cows from the pasture and supplements would have provided 20–55% of their daily P requirements of ~21 g P/d. Osteomalacia occurred in cattle at the 2 ranches as a result of severe P deficiency in the soil and forage, and inadequate P supplementation. Following diagnosis, control of P deficiency in beef cattle requires estimation of the amount of pasture P ingested and provision of sufficient additional supplementary P to meet the animals’ requirements.