Analysis of highbush blueberry (Vaccinium corymbosum L.) fruit growth with exponential mixed models

Blueberry fruit growth commonly exhibits a double-sigmoid pattern. The goal of this work was to characterize and compare fruit growth of cultivars differing in ripening time using exponential non-linear mixed models. Mixed-effects five-parameter exponential (Gompertz I and II; logistic; monomolecular) models were fitted to fruit diameter data from 2 years and three cultivars grown in the field in a cool temperate environment. Gompertz II mixed model provided the best fit to fruit growth data and was used for further analysis. In later ripening cultivars ‘Cape Fear’ and ‘Herbert’, clear-cut double-sigmoid patterns were observed, absolute growth rate models exhibited two marked peaks, and relative growth rate showed an initial decreasing trend, and a subsequent peak. The earlier cv. ‘O’Neal’ did not exhibit a defined double-sigmoid pattern. The time between relative extremes of absolute growth acceleration is proposed as an objective criterion for fixing stage boundaries within growth curves. Exponential mixed models accurately fitted blueberry growth patterns. These equations highlighted differences in fruit growth patterns between early and late ripening highbush blueberry cultivars.     

Studies on synchronous egress of coccidian parasites (<Emphasis Type="Italic">Neospora caninum,Toxoplasma gondii,Eimeria bovis)</Emphasis> from bovine endothelial host cells mediated by calcium ionophore A23187

Neospora caninum, Toxoplasma gondii and Eimeria bovis are coccidian parasites of veterinary importance. Tachyzoites of N. caninum and T. gondii and sporozoites of E. bovis are able to invade and replicate in endothelial cells in vivo and in vitro. As it holds true for all eukaryotic cells, the survival of parasitized host cells and the parasites themselves should be dependent on ion balances, especially on extra- and intracellular calcium concentrations. Addition of the calcium ionophore A23187 reliably did release merozoites from mature N. caninum and T. gondii meronts grown in cultured primary bovine umbilical vein endothelial cells (BUVEC). Extent and time course of merozoite release depended on both, maturity of the meronts and concentration of the calcium ionophore. Attempts, however, to achieve synchronous release of merozoites from E. bovis first generation meronts by ionophore treatment failed, suggesting a different biological behaviour of this parasite. According to microscopical observations, the quite variable time of E. bovis macromeront maturation and a hampered merozoite exit owing to dense parasite-induced cytoskeleton elements surrounding the meront may be a reason for the lack of inducible synchronous release. Electronic supplementary materials The online version of this article (doi: ) contains supplementary material, which is available to authorized users.     

Hypercalcemia associated with gastric pythiosis in a dog

A 20-month-old castrated male Labrador Retriever with a 3-month history of anorexia, weight loss, and vomiting was evaluated. Plasma biochemical abnormalities included marked hyperglobulinemia and hypercalcemia. Serum levels of parathyroid hormone, parathyroid hormone-related protein, 25-hydroxyvitamin D, and 1,25-dihydroxyvitamin D were either low or within reference intervals. Gastric wall thickening and abdominal lymphadenomegaly were observed with abdominal ultrasonography. Cytologic evaluation of a sample obtained via fine-needle aspiration of the gastric wall revealed pyogranulomatous inflammation and numerous poorly stained hyphae. Partial gastrectomy was performed, and a diagnosis of gastric pythiosis was made by immunohistochemical staining of infected gastric tissue, as well as by immunoblot serology. This case demonstrates that diagnostic samples for cytologic evaluation can be obtained by fine-needle aspiration of Pythium insidiosum-infected tissues and that a presumptive diagnosis can be made by examination of a Romanowsky-stained smear. Furthermore, pythiosis should be considered as a differential diagnosis for hypercalcemia, especially in young dogs with inflammatory lesions that have a granulomatous component. The mechanism for the hypercalcemia in this dog was not determined; however, calcium concentrations normalized after surgical resection of the gastric lesion.     

Assessing the efficacy of Duddingtonia flagrans chlamydospores per gram of faeces to control Haemonchus contortus larvae

The aims were (a) to quantify the number of Duddingtonia flagrans chlamydospores per gram of faeces (CPG) recovered from sheep administered with different oral doses and, (b) to describe the relationship between CPG and eggs per gram of faeces (EPG) on the efficacy to reduce Haemonchus contortus infective larvae. Three doses of chlamydospores per kg BW were orally administered during seven days: (T1) non treated control group, (T2) 1 × 10⁶, (T3) 2.5 × 10⁶ and (T4) 5 × 10⁶. Three lambs, infected with H. contortus, were used per group. Faeces were obtained from the rectum of each lamb during the fungal administration period (days 0–6) and for six days after that period. Four coproculture replicates were made from each animal in days 2, 4, 6, 8 and 10. A higher chlamydospore dose produced higher CPG in faeces (p < 0.05), but a clear dose dependent effect was not found either in the larvae reduction or in the CPG:EPG ratio. When ratios were re-analyzed, independently of the treatment groups of origin, a better efficacy was obtained with a ratio from 5 to 10 CPG:EPG and a higher ratio (>10 per egg) showed a lower reduction efficacy (p < 0.05). The binomial analysis showed that for each unit of increment in CPG:EPG ratio there was a reduction of larvae number until a point (between 5 and 10 CPG:EPG) where no further reduction was detected. The surface response test indicated that the number of larvae was reduced by CPG until possible saturation. The highest CPG:EPG ratios did not necessarily improve efficacy of D. flagrans.     

Rapid diagnosis and quantification of Francisella tularensis in organs of naturally infected common squirrel monkeys (Saimiri sciureus)

Francisella tularensis, a small Gram-negative facultative intracellular bacterium, is the causative agent of tularaemia, a severe zoonotic disease transmitted to humans mostly by vectors such as ticks, flies and mosquitoes. The disease is endemic in many parts of the northern hemisphere. Among animals, the most affected species belong to rodents and lagomorphs, in particular hares. However, in the recent years, many cases of tularaemia among small monkeys in zoos were reported. We have developed a real-time PCR that allows to quantify F. tularensis in tissue samples. Using this method, we identified the spleen and the kidney as the most heavily infected organ containing up to 400 F. tularensis bacteria per simian host cell in two common squirrel monkeys (Saimiri sciureus) from a zoo that died of tularaemia. In other organs such as the brain, F. tularensis was detected at much lower titres. The strain that caused the infection was identified as F. tularensis subsp. holarctica biovar I, which is susceptible to erythromycin. The high number of F. tularensis present in soft organs such as spleen, liver and kidney represents a high risk for persons handling such carcasses and explains the transmission of the disease to a pathologist during post-mortem analysis. Herein, we show that real-time PCR allows a reliable and rapid diagnosis of F. tularensis directly from tissue samples of infected animals, which is crucial in order to attempt accurate prophylactic measures, especially in cases where humans or other animals have been exposed to this highly contagious pathogen.     

An Ibero-American inter-laboratory trial to evaluate serological tests for the detection of anti-<Emphasis Type="Italic">Neospora caninum</Emphasis> antibodies in cattle

We carried out an inter-laboratory trial to compare the serological tests commonly used for the detection of specific Neospora caninum antibodies in cattle in Ibero-American countries. A total of eight laboratories participated from the following countries: Argentina (n = 4), Brazil (n = 1), Peru (n = 1), Mexico (n = 1), and Spain (n = 1). A blind panel of well-characterized cattle sera (n = 143) and sera representative of the target population (n = 351) was tested by seven in-house indirect fluorescent antibody tests (IFATs 1–7) and three enzyme-linked immunosorbent assays (ELISAs 1–3; two in-house and one commercial). Diagnostic performance of the serological tests was calculated and compared according to the following criteria: (1) the “Pre-test information,” which uses previous epidemiological and serological data; (2) the “Majority of tests,” which classifies a serum as positive or negative according to the results obtained by most tests evaluated. Unexpectedly, six tests showed either sensitivity (Se) or specificity (Sp) values lower than 90%. In contrast, the best tests in terms of Se, Sp, and area under the ROC curve (AUC) values were IFAT 1 and optimized ELISA 1 and ELISA 2. We evaluated a high number of IFATs, which are the most widely used tests in Ibero-America. The significant discordances observed among the tests regardless of the criteria employed hinder control programs and urge the use of a common test or with similar performances to either the optimized IFAT 1 and ELISAs 1 and 2.     

Supplementation of pig diets in the growth and termination phases with different calcium sources

The aim of this study was to evaluate the effect of supplementation of pig diets in the growth and termination phases with different calcium sources. In experiment I, 36 whole males were distributed in randomized blocks in six groups, with six replications. A basal diet was formulated to meet the animals’ nutritional requirements except for calcium (0.09%), and the sources evaluated (calcitic limestone, monodicalcium phosphate, calcinated bone flour, and oyster flour) replaced the basal diet to provide 0.59% of total calcium. To determine the endogenous calcium, a diet containing low calcium (0.019%) was given simultaneously to another group of animals. Feces and urine were collected for determination the coefficients of apparent and true digestibility. In experiment II, 160 piglets were distributed in randomized blocks in four treatments, with five replications and four animals per experimental unit. Carcass and performance parameters, calcium concentration in bone and serum, and bone parameters were evaluated. The data were submitted to analysis of variance and factorial. The calcium source did not influence the digestibility coefficients determined by total collection (P > 0.05). The digestibility of Ca from oyster flour estimated by collection with an indicator was higher than that from the other sources (P < 0.05). Calcium sources did not interfere in the evaluated parameters (P > 0.05). The sources studied in this work can be used to supplement growing pigs’ diets.     

Morphofunctional diversity of equine of varied genetic compositions raised in the Pantanal biome of Brazil

Evaluating phenotypic diversity makes it possible to identify discrepancies in aptitudes among animals of different genetic bases, which is an indicator of adaptive or selective differences between populations. The objective of this work was to evaluate the morphofunctional diversity of 452 male and female adult equines (Arabian, Quarter Mile, Pantaneiro, and Criollo breeds, and undefined crossbreeds of horses and mules) raised in the Pantanal biome (Brazil). Linear measurements were performed to estimate conformation indexes. Initially, a discriminant analysis was performed, regardless of the animal’s size, followed by factor analysis. The factors were characterized and used as new variables. The diversity among equines and their relationship with the factors were evaluated using multivariate analysis. The factors were classified according to their decreasing importance: balance, rusticity, and robustness for the measurement factors; and load, ability, conformation, and equilibrium for the index factors. The genetic groups of equines have well-defined morphofunctional characteristics. The main differences are based on the rusticity and ability typologies in relation to those based on performance. Equines introduced to the Pantanal biome presented a more robust and compact body with good conformation. As a result, these horses may have superior athletic performance during equestrian activities when compared to the Pantaneiro local breed. However, this biotype may represent less rusticity (less adaptive capacity). Therefore, the regional breed can be equal or better in equestrian activities than breeds introduced to the Pantanal biome. Thus, breeders may cross horses from local breeds as an alternative to those introduced. Undefined crossbred male equines presented a different profile from the Pantaneiro breed, which may indicate little use of crossbreeds in breeding.     

Keratitis due to microfilariae in dogs: a newly recognized disease

Parasitic agents have been associated with keratitis, but a diagnosis of parasitic keratitis has not been commonly made in domestic animals. The purpose of this study was to describe the clinical and histopathological findings in seven dogs with chronic keratitis caused by microfilariae diagnosed in Brazil. All dogs presented with superficial corneal opacities of varying degrees affecting the perilimbal and central regions of the cornea, with other opaque areas appearing as crystalline deposits and corneal vascularization. The lesions were bilateral and were associated with mild‐to‐moderate conjunctival hyperemia. There was no history of blepharospasm or pruritus, and no subjects presented with epithelial erosions. Corneal biopsy revealed free microfilariae in the corneal stroma, with varying degrees of inflammation and collagen fiber destruction. The microfilariae were also found in skin lesions by skin snip technique. No adult worms were found in these dogs, and no dogs were on heartworm preventative before diagnosis. Monthly doses of oral ivermectin improved ocular and dermal lesions. One dog showed complete remission with the treatment. The species of the microfilariae was not identified.     

Population pharmacokinetics of itraconazole solution after a single oral administration in captive lesser flamingos (Phoeniconaias minor)

Aspergillosis is an infectious, non‐contagious fungal disease of clinical importance in flamingo collections. Itraconazole is an antifungal drug commonly used in the treatment and prophylaxis of avian aspergillosis. Studies have shown that dosage regimes in birds vary based on different itraconazole presentation and administration methods. This investigation used a population pharmacokinetic approach to study itraconazole in lesser flamingos. Itraconazole was administered orally at 10 mg/kg to 17 flamingos. A sparse blood sampling was performed on the subjects, and samples were collected at 1, 2, 3, 5, 8, 12, 16, 21, and 24 hr post‐drug administration. Twelve flamingos were sampled three times, three birds bled twice and two sampled once. Itraconazole in plasma was quantified using high‐pressure liquid chromatography (HPLC). A one‐compartment pharmacokinetic model with first order absorption was fitted to the data using nonlinear mixed effects modeling (NLME) to determine values for population parameters. We identified a long half‐life (T½) of more than 75 hr and a maximum plasma concentration (C_MAX) of 1.69 µg/ml, which is above the minimal inhibitory concentrations for different aspergillus isolates. We concluded that plasma drug concentrations of itraconazole were maintained in a population of flamingos above 0.5 ug/ml for at least 24 hr after a single oral dose of 10 mg/kg of itraconazole solution.