Histochemical study of the anulus fibrosus in normal canine caudal cervical intervertebral discs

A qualitative assessment of glycosaminoglycans (GAGs) in the non-chondrodystrophoid canine cervical anulus fibrosus was performed using the Alcian blue/critical electrolyte concentration staining technique. Estimates of GAG concentration were deduced for two groups of dogs (average age four and eight years) by determining the hyaluronic acid, chondroitin sulphates and keratan sulphate 'alcianophilic indices'. Keratan sulphate was the predominant GAG in the four-year-old group of dogs. The concentration of keratan sulphate was equal to that of chondroitin sulphates in the eight-year-old group and the total GAG concentration was decreased. This qualitative assessment indicated that the concentrations of keratan sulphate decreased and chondroitin sulphates increased between four and eight years of age. Similar conclusions have not been reported for anuli in the thoracic and lumbar regions of the canine spine.     

Validation of the FAMACHA eye color chart for detecting clinical anemia in sheep and goats on farms in the southern United States

Recent studies on sheep and goat farms in the southern United States indicate that multiple-anthelmintic resistance in Haemonchus contortus is becoming a severe problem. Though many factors are involved in the evolution of resistance, the proportion of the parasite population under drug selection is believed to be the single most important factor influencing how rapidly resistance develops. Therefore, where prevention of resistance is an important parallel goal of worm control, it is recommended to leave a portion of the animals untreated. Recently, a novel system called FAMACHA was developed in South Africa, which enables clinical identification of anemic sheep and goats. When H. contortus is the primary parasitic pathogen, this system can be applied on the farm level to reduce the number of treatments administered, thereby increasing the proportion of the worm population in refugia. Since most studies validating the FAMACHA method have been performed in South Africa, it is important that the method be tested in other regions before its use is broadly recommended. We performed a validation study of FAMACHA by testing the system in sheep (n = 847) and goats (n = 537) of various breeds and ages from 39 farms located in Arkansas, Georgia, Louisiana, Florida, and the US Virgin Islands. The color of the ocular conjunctiva of all animals were scored on a 1-5 scale using the FAMACHA card, and blood samples were collected from each animal for determination of packed cell volume (PCV). Fecal samples were also collected from a majority of the animals tested for performance of fecal egg counts (FEC). Correlations between PCV and eye scores, PCV and FEC, and FEC and eye scores were all highly significant for both sheep and goats (P < 0.001). Data for both FAMACHA scores and PCV were evaluated using two separate criteria for anemia: eye score values of 3, 4 and 5 or 4 and 5, and PCV values of < or =19 or < or =15 were considered anemic. Specificity was maximized when eye score values of 4 and 5 were considered anemic and PCV cut off for anemia was < or =19, but sensitivity was low. In contrast, sensitivity was 100% for both sheep and goats when eye score values of 3, 4 and 5 were considered anemic and PCV cut off was < or =15, but specificity was low. In both sheep and goats, predictive value of a negative was greater than 92% for all anemia and eye score categories, and was greater than 99% for both eye score categories when an anemia cutoff of < or =15 was used. Predictive value of a positive test was low under all criteria indicating that many non-anemic animals would be treated using this system. However, compared to conventional dosing practices where all animals are treated, a large proportion of animals would still be left untreated. These data indicate that the FAMACHA method is an extremely useful tool for identifying anemic sheep and goats in the southern US and US Virgin Islands. However, further studies are required to determine optimal strategies for incorporating FAMACHA-based selective treatment protocols into integrated nematode control programs.     

α‐6 Integrin Expression in Bovine Spermatogonial Cells Purified by Discontinuous Percoll Density Gradient

The study of spermatogonial stem cells (SSCs) provides a model to better understand adult stem cell biology. Besides the biomedical potential to perform studies of infertility in many species, SSCs hold a promising application at animal transgenesis. Because stem cells are thought to be associated with basement membranes, expression of α‐6 integrin has been investigated as a marker of type A spermatogonial cells, which are considered SSCs because of their undifferentiated status and self‐renewal ability. In this manner, the aim of this study was to isolate type A SSCs from adult bulls by a two‐step enzymatic procedure followed by a discontinuous Percoll density gradient purification and verify the expression of α‐6 integrin by flow cytometry and real‐time RT‐PCR before and after Percoll purification. Spermatogonial cells were successfully obtained using the two‐step enzymatic digestion. An average of 1 × 10⁵ viable cells per gram of testis was isolated. However, the discontinuous Percoll did not purify isolated cells regarding α‐6 integrin expression. Flow cytometry analysis demonstrated no differences in the α‐6 integrin expression between cell samples before and after Percoll purification (p = 0.5636). The same was observed in the real‐time PCR analysis (p > 0.05). In addition to α‐6 integrin, the expression of GFRa‐1 and PGP9.5, known bovine SSCs markers, was detected in all samples studied. Considering that Percoll can reduce cell viability, it is possible to conclude that Percoll density gradient is not suitable to purify bovine SSC, according to α‐6 integrin expression.     

Use of a mixed sericea lespedeza and grass pasture system for control of gastrointestinal nematodes in lambs and kids

Because of a high prevalence of anthelmintic resistance and consumer demand for chemical free meat products, management tools to minimize the need for deworming are needed. The objective was to examine the effectiveness of grazing sericea lespedeza (SL) in a mixed grass or a pure forage system for control of gastrointestinal nematodes (GIN); in other words pasture systems included grass, grass plus SL, or SL alone (Experiments 2 and 3). Selective use of copper oxide wire particles (COWP) based on the FAMACHA(?) system was used to aid in GIN control. In Experiment 1, lambs co-grazed bermudagrass (BG; n=21) or SL in a mixed grass pasture (SLM; n=22) with dams for 14 days. In Experiment 2, lambs grazed BG (n=14), SLM (n=13), or pure SL (SLP; n=13) pastures for 56 days. In Experiment 3, doe kids grazed BG (n=12), SLM (n=13), or SLP (n=13) for 84 days. Animals were fed a 16% crude protein supplement based on NRC requirements and estimated forage quality of pastures, so that 454, 389, and 200 g/lamb (Experiment 2), or 454, 300, and 150 g of supplement/goat (Experiment 3) was fed to BG, SLM, and SLP, respectively. Animals were dewormed with COWP if FAMACHA(?) was >3. Coprocultures were conducted to identify GIN genus. In Experiment 1, FEC were reduced in lambs grazing SLM compared with BG pastures. In Experiment 2, FEC were reduced in SLP compared with BG lambs on all days, and reduced in SLM compared with BG lambs on day 56. Initially, Haemonchus contortus was the predominant nematode, but the population shifted to other species in the SL groups by the end of the study. The mean number of dewormings/lamb was 0.71, 0.20, and 0.21±0.13 for BG, SLM, and SLP groups, respectively. In goats in Experiment 3, Trichostrongylus spp. was the predominant nematode in May and June and H. contortus in July. There was little meaningful effect of forage treatments on GIN infection in kids. Because H. contortus was not the predominant nematode in kids, the integrated approaches used may not have been effective in controlling GIN. Grazing SL was effective for GIN control in lambs in early summer, but the effect was delayed in lambs grazing a mixed SL/grass pasture unless lambs initially grazed with dams. An integrated approach used that included SL grazing and COWP for deworming was effective in lambs.     

Hemodynamic effects of sevoflurane in spontaneously breathing cats

Sevoflurane has recently been introduced in feline anesthesia. However, its cardiovascular effects have not, to our knowledge, been reported in this species. Six healthy cats, aged 1.81 ± 0.31 years (mean ± SEM) and weighing 3.47 ± 0.11 kg, were studied. Anesthesia was induced and maintained with sevoflurane in oxygen. Body temperature was maintained between 38.5 and 39.55 °C. After instrumentation, end‐tidal sevoflurane concentration was randomly set at 1.25, 1.5, and 1.75 times the individual minimum alveolar concentration (MAC), determined in a previous study, according to a Latin Square Design. Thirty minutes of stabilization was allowed after each change of concentration. ECG and heart rate, systemic and pulmonary arterial pressures, central venous pressure (CVP), and core body temperature were continuously monitored and recorded. Inspired and end‐tidal oxygen, carbon dioxide, and sevoflurane concentrations were measured using a Raman spectrometer, calibrated every 80 minutes with three calibration gases of known sevoflurane concentration (1, 2, and 5%). Moreover, at selected times, pulmonary artery occlusion pressure and cardiac output (thermodilution) were measured, and arterial and mixed venous blood samples were collected for pH and blood gas analysis, hemoglobin concentration, hemoglobin oxygen saturation, packed cell volume (PCV) and total protein determination, and lactate concentration measurement. Cardiac index (CI), stroke index (SI), systemic and pulmonary vascular resistance indices, rate‐pressure product, left and right ventricular stroke work indices (LVSWI and RVSWI, respectively), arterial and mixed venous oxygen contents, oxygen delivery, oxygen consumption, and oxygen utilization ratio were calculated. Data were analyzed by a Repeated Measure Latin Square Design followed by a Tukey's test for 2 × 2 comparisons. Arterial pH significantly decreased from 7.40 ± 0.05 to 7.29 ± 0.07 with the administration of increasing concentrations of sevoflurane. Similarly, LVSWI decreased from 3.72 ± 0.60 to 2.60 ± 0.46 g m^?2. Mean arterial pressure, PaO₂, mixed venous pH, CI, SI, and oxygen delivery tended to decrease dose‐dependently, whereas CVP, PaCO₂, Pv CO₂, PCV, and arterial and mixed venous hemoglobin concentrations tended to increase dose‐dependently with the administration of sevoflurane. However, these trends did not reach statistical significance, possibly because of the limited number of animals studied. Sevoflurane seemed to induce dose‐dependent cardiovascular depression in cats.