猪败血型链球菌病的诊疗报告

本文阐述了猪链球菌病的临床症状、实验室检验及防治方法，力求为科学防控猪链球菌病提供有益的探索和参考。     

Effect of Several Antioxidants on Thawed Ram Spermatozoa Submitted to 37°C up to Four Hours

Thawed ram spermatozoa were incubated at 37°C in the presence of dehydroascorbic acid (DHA), TEMPOL (TPL), N‐acetyl‐cysteine (NAC) and rutin (RUT), at 0.1 and 1 mm , in order to test their effects on sperm physiology. Cryopreserved spermatozoa from four rams were thawed, pooled, washed and incubated in TALP‐Hepes with 1 mm or 0.1 mm of each antioxidant, performing a replicate with induced oxidative stress (Fe²⁺/ascorbate). Motility (CASA), viability and mitochondrial membrane potential (flow cytometry) were analysed at 2 and 4 h. Lipoperoxidation (MDA production), intracellular reactive oxygen species (ROS) and DNA status (TUNEL) were analysed at 4 h. Antioxidants, except DHA 0.1 mm , decreased motility and kinematic parameters, but had little effect on viability or mitochondrial activity. Except 1 mm DHA, the antioxidants reduced ROS at 4 h. Moreover, NAC 1 mm , rutin and TEMPOL reduced ROS and DNA damage in the presence of oxidative stress. N‐acetyl‐cysteine, rutin 1 mm and TEMPOL reduced lipoperoxidation in the presence of oxidative stress. However, DHA did not affect lipoperoxidation. At 1 mm , DHA increased DNA damage in the absence of oxidative stress. Dehydroascorbic acid effects could arise from spermatozoa having a low capacity for reducing it to ascorbic acid, and it may be tested in the presence of other antioxidants or reducing power. Future research should focus in testing whether the inhibition of motility observed for NAC, rutin and TEMPOL is reversible. These antioxidants might be useful at lower temperatures (refrigerated storage or cryopreservation) when their protective effects could be advantageous.     

Changes in Testicular Development, Ultrasonographic and Histological Appearance of the Testis in Buck Kids Immunized Against LHRH Using Recombinant LHRH Fusion Protein

This study was designed to evaluate the effectiveness of recombinant Ovalbumin-LHRL (OL) immunization on changes in testicular size, histological appearance and testosterone production in buck kids. Thirty native buck kids at 18 weeks of age were divided into three groups, control (n = 10), immunization (n = 10) and castration (n = 10) groups. Immunized animals received OL protein generated by recombinant DNA technology. Ultrasonographic and histological examinations of the testes were performed. Animals were slaughtered at 44 weeks of age. Semen and epididymides were evaluated for the presence of sperm cells. Immunized animals generated anti-LHRH antibodies. Testosterone production, testicular and accessory glands development and sperm production were suppressed in the immunized animals (p < 0.01). Semineferous tubule diameters decreased (p < 0.01), basal membrane of the tubule was thickened and hyalinized in immunized kids. Immunization affected ultrasonographic appearance of the testes drastically. While testes of control animals gained their normal ultrasonographic appearance as the age increased, immunized animals had uniform hypoechogenic testicular structure as observed at 18 weeks of age until slaughter. Simultaneous histological and ultrasonographic evaluations indicated that the changes in testicular histology could partly be monitored via ultrasonographic imaging; nevertheless, it is difficult to claim that ultrasonographic image reflects the exact changes in such instances. In conclusion, these results indicate that recombinant OL fusion protein is effective in immunocastration in buck kids and has a potential to be used as an alternative to physical castration. Further researches should be conducted to help assessing reproductive status of testes from ultrasound images.     

Refrigerated Storage of Red Deer Epididymal Spermatozoa in the Epididymis, Diluted and with Vitamin C Supplementation

We have approached the problem of refrigerated storage of epididymal sperm samples from red deer by comparing three options: storing the genital (testicles within the scrotum), diluting the semen in extender or diluting the semen in extender supplemented with an anti-oxidant. Twenty-nine pairs of testes were collected. Spermatozoa from one of each of the pairs were immediately recovered, and diluted to 400 × 10⁶ sperm/ml in Tris-citrate-fructose with 20% egg yolk. Control group was stored as such, and Anti-oxidant group was supplemented with 0.8 m m vitamin C. The remaining epididymides and the diluted samples were stored at 5°C and spermatozoa were analysed at 0, 24, 96 and 192 h for: motility [computer-assisted semen analysis (CASA)], acrosomal integrity, sperm viability (eosine/nigrosine staining), normal tails and chromatin status [sperm chromatin structure assay (SCSA)]. In general, seminal quality decreased with storage time. Vitamin C supported progressive motility better at 24 h (median 42% vs 23% Control and 15% epididymis), reduced the incidence of tail abnormalities and protected chromatin. Storing the semen in the epididymis slowed down motility loss, but slightly increased the occurrence of tail abnormalities and viability was lower at 192 h. However, regarding chromatin status, sperm stored in the epididymis was protected similarly to those diluted in the medium supplemented with vitamin C. Although the differences between the three groups were small, there were some advantages in supplementing the extender with vitamin C. Besides, refrigerating the epididymis may be a good option when immediate processing is not available.     

Reproductive Performance of Rabbit Does Artificially Inseminated via Intravaginal Administration of [des-Gly 10, d-Ala6]-LHRH Ethylamide as Ovulation Inductor

This study was aimed to evaluate the reproductive performance of rabbit does artificially inseminated (AI) with a GnRH analogue [des‐Gly10, d ‐Ala6]‐LHRH. ethylamide to induce ovulation by intravaginal administration, delivered in the seminal dose. In a preliminary experiment, 39 does were divided into three groups (n = 13) that, at the time of AI, received the following ovulation induction treatments: (i) control group: 20 μg of gonadorelin administered intramuscularly; (ii) 25 μg of the GnRH analogue added to the seminal dose; (iii) 30 μg of the GnRH analogue added to the seminal dose. Fertility did not differ between the three groups (control: 80.6%, group 2: 82.8%, group 3: 73.3%). In a second experiment, a large‐scale field trial was conducted to test the use of 25 μg of the GnRH analogue [des‐Gly10, d ‐Ala6]‐LHRH ethylamide delivered in the seminal dose (n = 270) against 20 μg of gonadorelin administered intramuscularly. Fertility was higher (p < 0.05) when ovulation was induced by intravaginal administration of the GnRH agonist (91.1% vs 85.6%). Prolificacy or mortality at birth was never affected by the ovulation induction treatments. In a third experiment, two groups of does [control group (n = 39): ovulation was induced using 20 μg of gonadorelin administered intramuscularly; treatment group (n = 40): ovulation was induced using 25 μg of [(des‐Gly10, d ‐Ala6)‐LHRH ethylamide added to the seminal dose] were inseminated at 42‐day intervals for five successive AI cycles, to test the response to the GnRH agonist after repeated intravaginal administration to the same animals. Fertility and prolificacy were not influenced by the ovulation induction treatment neither there was an interaction between treatment and parity. The last experiment was aimed to determine whether it could be possible to add the GnRH agonist to the semen in the AI Center, just after semen collection and dilution, or it would have to be added in the farm, immediately before AI. Kindling rates did not significantly differ when ovulation was induced by intramuscular injection of gonadorelin (84.5%) or when the GnRH agonist was added to the seminal dose just at the moment (93.8 %) or 24 h before AI (90.4 %), but it was significantly lower when the hormone was added to the semen 32 h before AI (76.3 %). Prolificacy, however, was not influenced by the ovulation induction treatment.     

Loss of Avirulence and Reduced Pathogenicity of a Gamma-Irradiated Mutant of Fusarium oxysporum f. sp. lycopersici

ABSTRACT The tomato Fusarium resistance gene I-2 confers resistance to F. oxy-sporum f. sp. lycopersici race 2, which expresses the corresponding aviru-lence gene avrI-2. To elucidate the molecular basis of this gene-for-gene interaction, we initiated a search for the avrI-2 gene. Gamma irradiation mutagenesis, using (137)Cs, was performed to generate an avrI-2 mutant of F. oxysporum f. sp. lycopersici. To this end, a race 2 isolate was first transformed with a phleomycine resistance gene and a GUS marker gene in order to distinguish mutants from contaminating isolates. A total of 21,712 mutagenized colonies was tested for loss of avirulence on I-2-containing tomato seedlings. One mutant was selected that showed the expected loss of avirulence but, surprisingly, also showed reduced pathogenicity toward susceptible tomato plants. DNA analysis was subsequently used to visualize genomic changes in the mutant. Southern analysis on contour-clamped homogeneous electrophoretic field blots demonstrated a translocation of a 3.75-Mb chromosome in the mutant. Random amplified polymorphic DNA and amplified fragment length polymorphism analysis identified at least nine polymorphisms between the wild-type and mutant isolates. Most of these polymorphisms appeared as extra fragments in the mutant and contained repetitive DNA sequences.     

Ram sperm survival during the use of various solvents after deep freezing in liquid nitrogen]

Suitability of three diluents for a deep-freezing of ram ejaculates and sperm survival after de-freezing and in the course of a six-hour incubation were investigated. The longest survival was found with the use of diluent containing Tris, fructose, and citric acid: 49.2% after de-freezing and 20.7% after six hours. The influence of the de-freezing procedure on the motility of spermatozoa can also be taken into consideration. The best proved rapid de-freezing without any diluent. The stated results serve for orientation only as the suitability of the diluents will be confirmed by the fertility of inseminated ewes.     

Coxiella burnetii Shedding During the Peripartum Period and Subsequent Fertility in Dairy Cattle

The objective of this study was to assess the effects of Coxiella burnetii shedding or seropositivity on post‐partum recovery and subsequent fertility in high‐producing dairy cows. Given the difficulty in diagnosing C. burnetii infection at the farm level, an exhaustive series of tests in 43 pregnant animals that delivered at least one live calf were conducted, including blood serology and PCR of milk or colostrum, cotyledons (only at parturition), faeces, vaginal fluid against C. burnetii on gestation Day 171–177, at parturition and on Days 1–7, 8–14, 15–21, 22–28, 29–35 and 90–97 post‐partum. During scheduled herd visits, ultrasonography (US) of the genital tract and examination of vaginal fluid were performed on Days 15–21 (V1), 22–28 (V2), 29–35 (V3) and 51–57 (V4) post‐partum by the same veterinarian. Logistic regression analysis revealed that the likelihood of suffering endometritis (the presence of echogenic intrauterine fluid (IUF), cervical diameter of ≥4 cm or endometrial thickness ≥0.75 cm) was lower in C. burnetii‐seropositive animals (OR = 0.10), compared with C. burnetii‐seronegative animals. According to Kaplan–Meier survival analysis, C. burnetii‐seronegative and non‐shedding cows showed a delayed return to luteal activity and conception was delayed in non‐shedding animals, compared with the remaining animals. Overall, the results of our study provide useful insight into the effects of C. burnetii infection on post‐partum recovery and subsequent fertility. In particular, animals not infected with Coxiella seem to be susceptible to infection and not protected against the bacterium in dairy herds. The elevated costs of determining an infection at the farm level, make monitoring of cows virtually impossible from a clinical point of view.