Estimation of S phase duration in goat epidermis by an in vivo intradermal double labeling technique using bromodeoxyuridine and tritiated thymidine.

When studying skin diseases resulting from alterations in the rate of epidermal cell turnover it is useful to be able to quantify parts of the epidermal cell cycle. An in vivo intradermal technique is described which uses tritiated thymidine followed by bromodeoxyuridine to label cells in the S phase of the cell cycle. Combined autoradiographic and immunocytochemical techniques were used to quantify the flux of cells into and out of S phase. These results were then used to estimate the length of S phase. The technique was found to provide clear distinctive labelling of S phase nuclei with both reagents. This avoided many of the problems encountered with double labelling techniques using two radioactive labels. S phase was calculated to be 7.7 hours for goat skin.     

Comparison of MTT colorimetric assay and tritiated thymidine uptake for lymphocyte proliferation assays using chicken splenocytes.

The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) colorimetric assay was compared with the conventional tritiated thymidine deoxyriboside (3H-TdR) incorporation for assay of lymphocyte blastogenesis using mononuclear cells isolated from the spleens of specific-pathogen-free chickens. The study was undertaken in an effort to simplify methods for assessing avian lymphocyte proliferation, specifically for evaluating response to mitogens or for indirect measurement of T-cell growth factors. The results from stimulated cells in both assay methods were significantly different from results from the control cells, and the MTT assay results regressed in a significant linear manner on counts from 3H-TdR incorporation. On this basis, the MTT assay is a valid test for evaluation of lymphocyte proliferation of chicken splenocytes.     

Comparison of invasive and oscillometric blood pressure measurement techniques in anesthetized sheep,goats, and cattle

ObjectiveTo determine the level of agreement between an oscillometric (O-NIBP) and an invasive method (IBP) of monitoring arterial blood pressure (ABP) in anesthetized sheep, goats, and cattle.Study designProspective clinical study.AnimalsTwenty sheep and goats, 20 cattle weighing <150 kg body weight, and 20 cattle weighing >150 kg body weight.MethodsAnimals were anesthetized and systolic ABP (SABP), mean ABP (MABP), and diastolic ABP (DABP) were measured using IBP and O-NIBP. Differences between IBP and O-NIBP, and 95% limits of agreement (LOA) between SABP, MABP, and DABP values were assessed by the Bland–Altman method.ResultsMean difference ± standard deviation (range) between SABP, DABP, and MABP measurements in sheep and goats was 0 ± 16 (-57 to 38) mmHg, 13 ± 16 (-37 to 70) mmHg, and 8 ± 13 (-34 to 54) mmHg, respectively. Mean difference between SABP, DABP, and MABP measurements in small cattle was 0 ± 19 (-37 to 37) mmHg, 6 ± 18 (-77 to 48) mmHg, and 4 ± 16 (-73 to 48) mmHg, respectively. Mean difference between SABP, DABP, and MABP measurements in large cattle was -18 ± 32 (-107 to 71) mmHg, 7 ± 29 (-112 to 63) mmHg, and -5 ± 28 (-110 to 60) mmHg, respectively. The 95% LOAs for SABP, DABP, and MABP were -31 to +31, -19 to +44, and -19 to +34 mmHg, respectively in sheep and goats; were -37 to +37, -19 to +44, and -19 to +34 mmHg, respectively in small cattle; and were -81 to +45, -50 to +63, and -59 to +50 mmHg, respectively in large cattle.ConclusionsAgreement was poor between O-NIBP and IBP monitoring techniques.Clinical relevanceArterial BP should be monitored in anesthetized sheep, goats, and cattle using IBP.     

Direct and indirect measurement of somatic cell count as indicator of intramammary infection in dairy goats

Background

Mastitis is the most important and costly disease in dairy goat production. Subclinical mastitis is common in goats and is mainly caused by contagious bacteria. Several methods to diagnose subclinical mastitis are available. In this study indirect measurement of somatic cell count (SCC) by California Mastitis Test (CMT) and direct measurement of SCC using a portable deLaval cell counter (DCC) are evaluated. Swedish goat farmers would primarily benefit from diagnostic methods that can be used at the farm. The purpose of the study was to evaluate SCC measured by CMT and DCC as possible markers for intramammary infection (IMI) in goats without clinical symptoms of mastitis. Moreover to see how well indirect measurement of SCC (CMT) corresponded to direct measurement of SCC (DCC).

Method

Udder half milk samples were collected once from dairy goats (n = 111), in five different farms in Northern and Central Sweden. Only clinically healthy animals were included in the study. All goats were in mid to late lactation at sampling. Milk samples were analyzed for SCC by CMT and DCC at the farm, and for bacterial growth at the laboratory.

Results

Intramammary infection, defined as growth of udder pathogens, was found in 39 (18%) of the milk samples. No growth was found in 180 (81%) samples while 3 (1%) samples were contaminated. The most frequently isolated bacterial species was coagulase negative staphylococci (CNS) (72% of all isolates), followed by Staphylococcus aureus (23% of all isolates). Somatic cell count measured by DCC was strongly (p = 0.000) associated with bacterial growth. There was also a very strong association between CMT and bacterial growth. CMT 1 was associated with freedom of IMI while CMT ≥2 was associated with IMI. Indirect measurement of SCC by CMT was well correlated with SCC measured by DCC.

Conclusions

According to the results, SCC measured with CMT or DCC can predict udder infection in goats, and CMT can be used as a predictor of the SCC.     

Experimental bovine Trichophyton verrucosum infection. Comparison of the rate of epidermal cell proliferation and keratinisation in non-infected and reinoculated cattle.

The rate of epidermal cell renewal in normal bovine skin and that reinoculated with Trichophyton verrucosum was measured using a radioautographic technique. The transit times of both nucleated and fully keratinised cells were measured in sequential biopsy samples removed at predetermined periods after intradermal inoculation with radio-labelled isotopes. The total time taken for cells of the basal layer to travel to the point of desquamation in the stratum corneum was 18 days in normal cattle. In similar areas on cattle that had been reinoculated with T verrucosum the total epidermal cell renewal time was reduced to 12 days. Increased protein synthesis, as measured by incorporation of radio-labelled nucleoside was evident in basal cells within 24 h of reinoculation with the fungus. The nucleated epidermal cell thickness had almost doubled in areas of reinoculated skin within 72 h and increased cell proliferation was maintained for at least 10 days. Desquamation of the thickened stratum corneum had occurred within seven days of reinoculation with the fungus.     

两种片形吸虫感染山羊的对比试验

片形吸虫病是在全球范围广泛流行，并对人有一定危害的寄生虫病。在大部分地区该病病原为肝片形吸虫（Fasciola hepatica），非洲、亚洲的热带地区是大片形吸虫（F．gigantica）。据本地的资料和以往的研究显示，广西主要是大片形吸虫。由于国内外对肝片形吸虫的研究较多，对大片形吸虫的研究较少，使得国内一些研究未能注意到这两个种的虫在对动物的致病性及其所引起的免疫反应等方面的差别。     

In vivo measurement of bone strain in the horse.

Strain gauges were successfully bonded in vivo to the cranial, caudal, medial, and lateral aspects of the equine radium and tibia and to the dorsal, palmar, or plantar, medial, the lateral aspects of the metacarpus and metatarsus--all in the mid-diaphyseal region. Various activities were investigated, including walking, trotting or pacing, and standing up from anesthesia. The strain patterns showed that each stride produced a characteristic deformation cycle. The strains were measured and the axial loads were calculated as the horse performed certain activities. The tension band side of each bone was predicted from the results. The tension band sides of the metacarpus and metatarsus were the dorsomedial and dorsolateral aspects; for the radius and tibia, the tension band sides were the cranial and craniolateral aspects, respectively.     

Comparison between IMMUNODOT tests and the intradermal skin test in atopic dogs

This study evaluated a new perspective in the diagnosis of dermatitis in dogs with signs suggestive of allergic skin disease. The results obtained with CMG IMMUNODOT tests using the technique of allergen-specific strip tests, as employed for human allergy diagnosis, were compared with those obtained by the intradermal skin test (IDST). Forty-eight cases completed the diagnostic evaluation, which included IDST, flea-control program, exclusion of sarcoptes and, for some cases, a 1- to 2-month stabilization period on a restricted protein source diet and testing the serum in the presence of allergen-specific IgE and total IgE. The most common disorders included house and storage dust mites, allergic dermatitis and flea-allergic dermatitis together with atopy. This was confirmed serologically. In the case of positive IDST to pollens, Aspergillus spp. and cat epithelium, CMG IMMUNODOT strip tests were negative. A total of 25% of cases were considered to be primarily associated with food hypersensitivity, but only 4% were confirmed serologically. This study emphasizes the value of CMG IMMUNODOT tests as a support in the diagnosis of dog allergy.     

Comparison of serum cortisol concentration before and after intradermal testing in sedated and nonsedated dogs.

Serum cortisol concentration was evaluated in 71 dogs before and after a stressful procedure was performed. Thirty dogs were skin tested with sedation (group S), 21 dogs were skin tested without sedation (group NS), and 20 dogs had other dermatologic procedures performed (group C). Group-S dogs had significant (P less than 0.001) decrease in serum cortisol concentration after skin testing, compared with baseline values. In contrast, dogs of groups NS and C had significant (P less than 0.001) increase in poststress serum cortisol concentration. Mean cortisol concentration after stress was significantly lower for dogs of group S, compared with that for dogs of the other 2 groups. The second part of the analysis consisted of determining the number of false-negative skin test results for dogs of groups S and NS and comparing these with serum cortisol concentration. Difference in the number of suspected atopic dogs with negative skin test results (false-negative) was not evident between groups S and NS. Also, difference was not apparent between cortisol concentration in dogs that had positive or false-negative skin test results in either group. This finding indicates that high serum cortisol concentration does not affect results of skin testing in suspected atopic dogs.     

Incorporation of tritiated thymidine, leucine and histidine in murine tail epidermis after skin irritation (histoautoradiography).

Thymidine incorporation into epidermal DNA as well as leucine and histidine incorporation into epidermal protein were studied by histoautoradiography in female mice tail epidermis. Prior to in vitro incubation of skin samples with labelled precursors, tail skin was irritated mechanically by rubbing with fine sand paper, chemically by repeated topical administration of n-hexadecane and by feeding an essential fatty acid deficient diet. After these skin irritations, an increased thymidine labelling index and skin thickening were found. Leucine was shown to be incorporated into protein predominantly in basal epidermal cell layers, while histidine was incorporated predominantly in the granular layer. Especially after mechanical skin irritation, this difference was obvious.     

不同品种绵羊和山羊主要先天性免疫指标的比较

对4个山羊品种和4个绵羊品种的7项主要先天性免疫学指标进行了检测并比较。对动物进行颈静脉采血,制备血清,用ELISA试剂盒测定IL-1、IL-2、IL-6、IL-18、IFN-γ、NK细胞、血清溶菌酶。利用生物统计学的方法和原理,以品种和品种来源地为应变量对检测结果进行比较分析。结果表明,山羊当地品种的NK和LYS高于引进品种（P〈0.05）;绵羊引进品种的IL-1、IL-2、IL-6、IL-18、IFN-γ、NK、LYS均明显高于地方品种（P〈0.01）。在不同品种之间,莱芜黑山羊NK和LYS水平高于崂山奶山羊、波尔山羊和鲁波山羊,萨福克羊的IL-1、IL-6、IL-18、IFN-γ、LYS水平最高且差异极显著（P〈0.01）。小尾寒羊IL-18、IFN-γ水平最低且差异极显著（P〈0.01）。说明不同种群绵羊、山羊的先天性免疫水平存在显著差异,为以后抗病育种提供合理的试验数据。     

Response of cows with lymphoma to the intradermal injection of tumor cell antigens and phytohemagglutinin.

Tumor cell-membrane antigens did not elicit specific intradermal reactions in cows with lymphoma and were therefore not effective in identifying animals with tumor. Sixty-two percent (8/13) of cows with lymphoma in the advanced stages of disease responded poorly to the intradermal injection of a nonspecific mitogen (PHA-M) and normal lymphoid cell extracts. This finding may explain the lack of response to the tumor cell extracts that some cows with lymphoma and indicates the presence of immune deficits in animals with lymphoma. Six of eight animals with tumor and poor cutaneous sensitivity had sera that inhibited the blastogenesis of normal lymphocytes. The presence of this inhibitory material is associated with the presence of tumor, not with bovine leukemia virus infection and partially accounts for the relative cutaneous anergy in cattle with lymphoma.     

Comparison of absorbable and nonabsorbable sutures for intradermal skin closure in cats

Paired skin incisions were made in 6 cats and closed intradermally with the copolymer of glycolide, ɛ-caprolacton, and trimethylene-carbonate, or polypropylene suture. The macroscopic and histologic appearance of the incisions was compared. Polypropylene suture compared favorably to glycolide, ɛ-caprolacton, and trimethylene-carbonate suture for closure of skin incisions in cats.     

The cell count in milk of goats and the diagnosis of mastitis in goats (author's transl)

The cell counts in milk of goats seem to vary quite a lot. It is difficult to diagnose mastitis in goats using only cell counts as a criterion. Clinical symptoms correlated with bacteriology and differences in the cell counts of milk samples from the two halves are probably the criteria of choice in the laboratory mastitis diagnosis in goats.     

Dermal cellular responses of helminth-free and Ostertagia ostertagi-infected calves to intradermal injections of soluble extracts from O. ostertagi L3 larvae

Twelve calves were raised helminth-free until 9 weeks of age when six were orally inoculated with 100,000 Ostertagia ostertagi infective stage larvae (L3). Three uninfected and three experimentally infected calves received intradermal injections of sterile saline and soluble larval extract (SLE) from O. ostertagi L3 with a protein concentration ranging from 1 to 200 micrograms ml-1. Biopsies were performed 48 h post-injection. A kinetic study was performed on the remaining six calves, three infected and three uninfected, using a 100 micrograms ml-1 concentration of SLE and taking biopsies 1, 4, 8, 12, 24, and 72 h post-injection at both the saline and SLE-injected sites. All calves had an immediate wheal and increase in skin thickness at the SLE-injected sites. The numbers of eosinophils infiltrating SLE-injected sites as compared to saline-injected sites were significant in both uninfected and infected calves, but the infected calves had significant numbers to a wider range of SLE concentrations and had significantly higher numbers than uninfected calves in the kinetic study. Infected calves also had significant numbers of basophils in the dose response study at concentrations of 5 and 100 micrograms ml-1 SLE. Neutrophil infiltration was similar in both groups and was significant at SLE-injected sites early in the kinetic study. Detectable mast cells were decreased in SLE-injected sites of infected animals and perivascular accumulation of mononuclear and some polymorphonuclear cells was observed in the deep dermis of infected animals.     

Comparison of pepsinogen forms in cattle, sheep and goats

Different types and subtypes of pepsinogen extracted from bovine, ovine and caprine abomasa were found to differ according to their phosphate content and relative molecular mass (Mr). Bovine pepsinogens had organic phosphate contents ranging from 1.65 to 2.22 mol of phosphate mol-1 of pepsinogen. Ovine pepsinogens were in the range 1.50 to 2.36 and caprine pepsinogens were in the range 1.42 to 2.00. The major types of pepsinogen from each species were different in size. Bovine pepsinogen had an Mr of 39,000, ovine had an Mr of 43,000 and caprine pepsinogen had an Mr of 42,000.     

Comparison between a gamma-IFN assay and intradermal tuberculin test for the diagnosis of bovine tuberculosis in field trials in Brazil.

Bovine tuberculosis is a major problem in Brazil. The intradermal tuberculin test is the standard test for detection of bovine tuberculosis in Brazil but can lack both sensitivity and specificity. The purpose of this study was to compare a bovine gamma-IFN assay with the tuberculin test under field conditions in Brazil. A total of 1632 animals from 13 dairy farms were tested using the single cervical tuberculin test (SCTT). Among those animals, about 15% of each herd, 220 in total, represented a high-risk group and were selected to be tested using the gamma-IFN test. Of the 1632 animals tested, 207 presented significant reactions representing 12.7% of the cattle studied. In the selected group the number of animals positive by the gamma-IFN assay was 126/220 (57.3%) and the total number of reactive cows on SCTT was 106/220 (48.2%). The real number of infected cattle, following standards, was 120/220 (54.5%). From these results the relative sensitivity rate of gamma-IFN test was 100% including the false-positive results and 88.3% for the SCTT--a significant (P < 0.01) difference in favour of the gamma-IFN test of 11.7%. The gamma-IFN assay also identified some positive animals 60-120 days earlier than the SCTT. In conclusion, we believe that the gamma-IFN assay can be used alone or in combination with the SCTT, as a valuable tool for the control of bovine tuberculosis in the Brazilian national herd.