兔精液冷冻复温后超微结构变化和低渗膨胀实验

实验从精子超微结构的变化对冷冻解冻精子的损伤进行研究。结果表明冷冻复温后兔的一些精子不同程度出现了质膜膨胀、变薄、皱褶及损伤；顶体肿胀、顶体外膜、内膜囊泡化或不连续，及顶体完全脱落；部分精子中段质膜破损。线粒体裸露、断裂、电子密度降低或部分丢失；少数主段末段质膜破损，抽丝断裂、散开。实验验证HOST（hypoosmotic swelling test)适合于兔精子膜功能活性检测。     

A Systematic Review of Studies Performing the Hypo‐Osmotic Swelling Test to Evaluate the Quality of Canine Spermatozoa

The hypo‐osmotic swelling test (HOS test) is a simple and inexpensive test to evaluate the functional integrity of sperm cell membranes. According to the existing literature, its simple applicability has turned it into a valuable additional parameter to standard canine semen analysis. In the recent years, much research has been conducted in this field. The aim of this systematic review was to evaluate the quality of published literature in canine reproduction concerning the HOS test. Using two distinguished databases, 38 articles were detected and analysed subsequently according to various aspects, for example study design, population, semen sampling and implementation concerning the HOS test. Although there are numerous articles available, the diagnostic value of the HOS test remains ambiguous. Until now, neither a recognized test protocol nor reliable reference values have been defined. Most of the trials evaluated show serious methodological flaws and therefore do not permit drawing reliable conclusions. According to our results, approximately half of the studies (n = 20) included a sample size of five or less animals. None of the studies examined the inter‐ or intraobserver agreement for the HOS test. Further research is warranted including appropriate statistical methods and a sufficient number of animals to establish a standardized test protocol as well as reliable reference values. Most importantly, it is required to clarify a correlation between the HOS test and the fertilizing capacity to determine the diagnostic value of the HOS test.     

Effects of Incubation Temperature and Semen Pooling on the Viability of Fresh,Chilled and Freeze‐Thawed Canine Semen Samples

This study assessed the effects of different incubation temperatures on semen viability and the influence of pooling on semen longevity. In experiment 1, semen samples were collected from five dogs, individually processed (individual semen: IS) and then aliquots from each male were pooled (pooled semen: PS). Semen samples (IS and PS) were diluted in a Tris‐glucose‐yolk extender and preserved as fresh (37 and 25°C) and chilled semen (4°C). Sperm motility and the percentages of sperm abnormalities and acrosome membrane integrity were assessed for 24 h. Storage at 25 or 4°C for the first 24 h yielded similar semen quality, but incubation at 37°C caused drastic reduction in sperm motility from 8 h of incubation onwards. In experiment 2, the semen was processed in the same way to that of experiment 1 and then preserved at 25 or 4°C until semen inactivation. Semen that was incubated at 25°C became completely inactive after 3–4 days of storage, while semen that was preserved at 4°C presented with more gradually decreased sperm motility (mean values of 40–60% for the first 8 days). In addition, the mixing of semen was only observed to influence the sperm quality of the samples stored at 4°C. In experiment 3, semen was collected from five dogs, pooled and frozen in liquid nitrogen; after thawing, it was preserved at 37, 25, 15 and 4°C, and the sperm quality was defined. The motility of the freeze‐thawed semen samples decreased quickly in the first 4 h after thawing, regardless of the preservation temperature of the thawed semen. This study confirmed that semen preserved at 37°C should be used within a maximum of 12 h, while the semen stored at 25°C shows acceptable quality for 24 h. Chilled semen presented highest most sustainable quality, especially when semen is processed as pooled semen.     

Canine Bladder and Urethral Tumors: A Retrospective Study of 115 Cases (1980–1985)

One hundred and fifteen dogs with neoplasms of the lower urinary tract (bladder and/or urethra) were retrospectively evaluated at five referral institutions participating in ongoing studies by the Veterinary Cooperative Oncology Group. Most tumors were malignant (97%) and of epithelial origin (97%). Lower urinary tract tumors were more common in older dogs weighing greater than 10 kg. The following significant (P less than 0.05) statistical associations were found using the University of Guelph hospital population as control; there was no sex predisposition although the female:male ratio was 1.95:1. Neutered dogs were predisposed as were Airedale Terriers, Beagles, and Scottish Terriers, whereas German Shepherds were significantly under-represented among dogs with lower urinary tract tumors. These statistical associations should be interpreted cautiously because of possible demographic differences in hospital populations among the University of Guelph and other cooperating institutions. There were no significant correlations between age, gender, weight, breed, response to therapy, and survival time. Clinical signs were indicative of lower urinary tract disease and included hematuria, stranguria, and pollakiuria. The laboratory data were nonspecific except for urinalysis test results. Hematuria and inflammatory urinary sediments were most commonly reported; neoplastic cells were identified in the urine sediment of 30% of dogs with lower urinary tract tumors. Contrast cystography was a useful noninvasive diagnostic method since 96% of the dogs had a mass or filling defect in the lower urinary tract demonstrated by this technique.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of N‐acetyl‐L‐cysteine Supplementation in Semen Extenders on Semen Quality and Reactive Oxygen Species of Chilled Canine Spermatozoa

The objective of this study was to evaluate the quality of chilled dog semen processed with extenders containing various concentrations of N‐acetyl‐L‐cysteine (NAC). Ejaculates from five dogs were collected, pooled and evaluated for concentration, motility, rapid steady forward movement (RSF‐movement), viability, acrosomal integrity and by the hypo‐osmotic swelling test (HOST). In addition, superoxide anion (O₂^‐?) production, hydroxyl radicals (OH^?) and total reactive oxygen species (tROS) were determined. The pool was divided into five aliquots, which were diluted to a final concentration of 66.66 × 10⁶ spermatozoa/ml with Tris‐glucose‐egg yolk extender containing one of the following concentrations of NAC (0, 0.5, 1, 2.5 or 5 mm ). The semen aliquots were chilled and preserved at 4°C. Semen quality was evaluated after rewarming at 72 h. Sperm motility was significantly higher with the 0.5 mm concentration compared with the control group (p = 0.001). Rapid steady forward movement was higher with the 0.5 and 1 mm concentrations compared with the control and 5 mm group (p < 0.001). Viability and HOST percentages were not significantly altered. Compared with the control, the 5 mm concentration showed significantly reduced percentages of spermatozoa with normal acrosomes (p = 0.049). None of the ROS values at 72 h were significantly affected by the presence of NAC in semen extenders, although all NAC concentrations showed lower O₂^‐? and OH^? values compared with the control. Only the concentrations of 1 and 5 mm inhibited the significant increase of tROS values after 72 h, compared with the fresh semen value. In conclusion, NAC supplementation of semen extenders is beneficial to semen motility of canine spermatozoa during chilling with the 0.5 mm concentration being the most effective, although no significant ROS inhibition was observed at 72 h.     

Effect of Albumin and Polyvinyl Alcohol on the Vitality, Motility and Acrosomal Integrity of Canine Spermatozoa Incubated in vitro

Sperm culture media used for in vitro fertilization (IVF) procedures are important factors concerning the viability, motility and acrosomal integrity of spermatozoa. The aim of this study was to investigate the effects of three different sperm diluting media, tissue culture medium (TCM‐199), sperm culture medium (Sp‐TALP) and human tubular fluid (HTF) supplemented with varying concentrations of bovine serum albumin (1, 4 and 6%) or polyvinyl alcohol (0.8%) on the acrosomal integrity, motility and viability of canine spermatozoa. Ejaculates collected from four dogs were diluted in all media and spermatozoa were separated from seminal plasma by the swim‐up technique. Sperm progressive motility was assessed using a phase contrast microscope. Viability and acrosomal integrity were evaluated using a dual stain technique (Giemsa–Trypan blue). The results demonstrated that the number of live canine spermatozoa was similar in culture media supplemented or not supplemented with macromolecules. A minimal concentration of albumin (1%) in the three media showed similar effects on vitality, motility and acrosomal integrity, as had higher concentrations (4 and 6%). The percentage of acrosome‐intact spermatozoa was markedly higher after HTF (94.1%) than after TCM‐199 (70.1%) or Sp‐TALP (71.0%) without supplementation. It is concluded that serum bovine albumin, irrespective of the concentration, preserved sperm viability and function, and HTF is the most suitable medium for preserving the acrosome in canine spermatozoa prepared for in vitro manipulation through short incubation.     

Immunization of Pups with Maternally Derived Antibodies to Canine Parvovirus (CPV) Using a Modified‐Live Variant(CPV‐2b)

The results of vaccination trials carried out on pups with maternally derived antibodies (MDA) to canine parvovirus (CPV), using a modified‐live CPV‐2b variant vaccine (29‐97/40 strain), are reported. The vaccine was able to overcome the obstacle of MDA, and to elicit protective immunity in 100% of the pups whose antibody titres were 1:10–1:40, 83% of the pups with titres of 1:80, 57% of the pups with titres of 1:160, and even in 60% of the pups with antibody titres of 1:320.     

Expression of Matrix Metalloproteinases (MMP‐2, MMP‐9) and their Inhibitors (TIMP‐1, TIMP‐2) in Canine Testis,Epididymis and Semen

This study aims to investigate the role of matrix metalloproteinases (MMPs) in determining semen quality and to evaluate the expression and cellular localization of MMP‐2, MMP‐9, tissue inhibitor of metalloproteinase‐1 (TIMP‐1) and TIMP‐2 in the testes, epididymis and ejaculated spermatozoa. Gelatinase activities between normal (n = 21) and abnormal (n = 25) semen samples showed a significant, sixfold increase in proMMP‐2 and MMP‐2 activity in high than low sperm concentration samples (p < 0.001). ProMMP‐9 and MMP‐9 levels were significantly elevated in samples with low sperm counts compared to those with high sperm density (p < 0.001). High levels of proMMP‐2 and MMP‐2 were associated with high sperm motility (≥70%, p < 0.001). Sperm‐rich fraction showed significantly (eight‐fold) higher proMMP‐9 enzymatic activity compared with prostatic fraction. The mRNA expressions of MMP‐2, MMP‐9, TIMP‐1 and TIMP‐2 were confirmed in testicular and epididymal tissues. Immunohistochemical staining illustrated the MMP‐2‐specific strong immunoreactivity in the head of mature spermatids during spermatogenesis, whereas MMP‐9, TIMP‐1 and TIMP‐2 were absent in these cells. Matrix metalloproteinase‐9 immunoreactivity was observed in the spermatocyte and round spermatid, whereas TIMP‐1 was only exhibited in the residual bodies. Immunolabeling of epididymal and ejaculated sperm demonstrated MMP‐2 localization along acrosomal region of sperm, while MMP‐9, TIMP‐1 and TIMP‐2 localization was merely limited to the flagella. In conclusion, spermatozoa initially acquire MMP‐2 during their formation at testicular level, and the presence of this protein persists through the epididymal transit and up to ejaculate. The enzymatic activity of MMP‐2 and MMP‐9 may serve as an alternative biomarker in determining semen quality.     

The use of Oral Milbemycin Oxime (Interceptor®) in the Treatment of Chronic Generalized Canine Demodicosis*

Abstract— The purpose of this clinical study was to evaluate the efficacy of orally administered milbemycin oxime (Interceptor®, Ciba-Geigy, Greensboro, U.S.A.) in the treatment of 46 dogs with chronic generalized demodicosis. Two dosage protocols were followed with milbemycin oxime dosed at 0.5–2.3 mg.kg^-1 once daily (low dose) or twice daily (high dose). Seventy six per cent of the dogs on the low dose protocol achieved negative skin scrapings (remission) within 90 days, 24% were failures, and 19% remain negative with 1 year follow-ups. Seventy five per cent of the patients that achieved remission on the low dose protocol relapsed within 1 year when the milbemycin was discontinued. Ninety six per cent of the dogs on the high dose protocol achieved negative skin scrapings within 90 days, 4% were failures, and 42% remain negative with 1 year follow-ups. Twenty nine per cent of the patients that achieved remission on the high dose protocol relapsed within 1 year when the milbemycin was discontinued. Résumé— Le but de cette étude clinique était d'évaluer l'efficacité de la mylbémycine par voie orale (Interceptor®, Ciba Geigy, Greensboro, U.S.A.) dans le traitement de 46 chiens atteints de démodécie généralisée. Deux posologies ont été utilisées: 0.5–2.3 mg.kg^-1 une fois par jour (dose faible) et deux fois par jour (dose forte). Soixante dix ourcent des chiens ayant subi le protocole faible dose ont présenté des raclages cutanés négatifs (rémission) au bout de 90 jours, 24% n'ont jamais répondu et 19%étaient toujours négatifs un an après le traitement. Soixante dix pourcent des animaux ayant répondu à ce traitement ont rechuté dans l'année suivant l'arrêt de la milbémycine. Quatre vingt dix pourcent des chiens ayant subi le protocole à dose forte ont présenté des raclages négatifs au bout de 90 jours, 1% n'ent jamais guéri et 42% sont restés négatifs un en après l'arrêt du traitement. Vingt neuf pourcent des patients ayant guéri à cette posologie ont rechuté dans l'année suivant l'arrêt de la mylbémycine. Zusammenfassung— Das Ziel der vorliegenden Untersuchung war die Ermittlung der Wirksamkeit von oral angewendetem Milbemycinoxim (Interceptor®, Ciba-Geigy, Greensboro, USA) bei der Behandlung von 46 Hunden mit chronischer generalisierter Demodikose. Bei der Dosierung von Milbemycinoxin wurden zwei Dosierungsprotokolle eingesetzt, bei der niedrigen. Dosierung wurde 0.5–2.3 mg/kg einmal täglich, bei der hohen zweimal täglich verabreicht. 76% der Humde mit der niedrigen Dosierung zeigten negative Hautgeschabsel (Remission) innerhalb von 90 Tagen, 24% sprachen nicht an. Bei 19% der Hunde blieben die Geschabsel negativ bei den Kontrolluntersuchungen über 1 Jahr Dauer. Bei 75% der Patienten, die mit der niedrigen Dosierung zur Remission gebracht wurden, traten Rückfälle innerhalb von 1 Jahr nach Absetzen des Milbemycin auf Bei 96% der Hunde aus der Gruppe mit der hohen Dosierung waren die Hautgeschabsel innerhalb von 90 Tagen negativ, 4% sprachen nicht auf die Behandlung an. Bei 42% der Tiere blieben die Geschabsel negativ bei den Kontrolluntersuchungen über 1 Jahr Dauer. Bei 29% der-Patienten, die unter der hohen Dosierung zur Remission gebracht waren, traten Rückfälle innerhalb von 1 Jahr nach Absetzen des Milbemycins auf. Resumen El propósito del siguiente estudio clínico fué la evaluación de la eficacia de la administración oral de óximo de milbecina, (Interceptor®, Ciba-Geigy, Greensboro, U.S.A.) en el tratamiento de 46 perros con demodicosis crónica generalizada. Los protocolos de dosificación con óximo de milbezina fueron los siguientes: 0.5–2.3 mg/kg una vez al día, (dosis baja), o dos veces (dosis alta). Setenta y seis por ciento de los perros que siguieron el protocolo a la dosis menor presentaron raspados cutáneos negativos, (remisión), en noventa dias, el 24% resulto en fracaso y el diecinueve por ciento permaneció negativo después de un año. El 75% de los pacientes que alcazaron la remisión a la dosis baja sufrieron un relapso al año de tratamiento cuando fue discontinuado. Noventa y seis de los perros con el protocolo de la dosis alta presentaron raspados cutáneos negativos a los noventa dias, el 4% fue un fracaso y el 42% permaneció con resultados negativos despues de un año. Veintinueve por ciento de los pacientes que alcanzaron la remisión a la dosis mayor presentaron relapso después de un año cuando el tratamiento fue discontinuado.     

Retrospective evaluation of the use of amiodarone in dogs with arrhythmias (from 2003 to 2010)

Objectives : To evaluate the efficacy of amiodarone in dogs with refractory supraventricular and ventricular arrhythmias and to document the side effects in treated dogs. Methods : Records of 28 dogs were retrospectively searched to document indication for amiodarone administration, heart rate, alkaline phosphatase, alanine aminotransferase, thyroxine (T4) and thyroid stimulating hormone values before and after starting treatment and during follow‐up periods. Results : Sixteen dogs with supraventricular and 12 dogs with ventricular arrhythmias were treated with amiodarone. Amiodarone treatment significantly reduced the heart rate (P<0.001) and resulted in improvement in the severity of the arrhythmia and clinical signs in 26 dogs. There were no significant differences in alkaline phosphatase (P=0.596), alanine aminotransferase (P=0.842), T4 (P=0.789) and thyroid stimulating hormone (P=0.064) before and after starting amiodarone. On maintenance therapy, median amiodarone blood levels were within the accepted reference range (0.5 to 2.0 mg/L) at 0.8 mg/L (range 0.2 to 11.6 mg/L), but the majority of the desethylamiodarone levels were below normal at 0.1 mg/L (range 0.1 to 0.9 mg/L), based on human reference intervals (0.5 to 2.0 mg/L). Clinical Significance : Amiodarone may be an effective and safe alternative to treat supraventricular and ventricular arrhythmias in dogs, when common anti‐arrhythmic drugs are not effective or contraindicated.     

Influence of Temperature during Glycerol Addition and Post-thaw Dilution on the Quality of Canine Frozen Semen

Contents The aim of the present study was to compare the influence of room temperature (27 degrees C) and 4 degrees C during glycerol addition on canine semen cryopreservation and verify the effect of different post-thawing dilutions on canine semen. Ten ejaculates from five stud dogs were collected by digital manipulation. Semen samples were evaluated and further divided into two aliquots. The first aliquot was extended in Tris-egg yolk-glycerol at 27 degrees C and the second one received glycerol at 4 degrees C. Samples were frozen and stored in liquid nitrogen. After 1 week, samples were thawed and submitted to evaluations of progressive sperm motility, morphology, acrosomal integrity, hypo-osmotic swelling (HOST) and thermoresistance tests. For thermoresistance test, aliquots were divided in two portions: one portion was kept undiluted (1 : 0) and the other one was diluted in a 1 : 4 ratio (one part semen to four parts extender). No differences were observed between temperatures for glycerol addition regarding seminal parameters evaluated. Furthermore, post-thawing dilutions demonstrated similar effect on canine semen longevity. Correlations among post-thaw sperm motility and HOST and results from thermoresistance test were observed for both temperatures for glycerol addition. In conclusion, glycerol could be added to canine semen at room temperature (27 degrees C) or at 4 degrees C. Moreover, there is no need to extend canine semen after thawing for the thermoresistance test, but if we need to increase the inseminating volume for artificial inseminations, the addition of extender will not damage the semen.     

19种杀虫剂对桑蚕毒性的测定与评价

本文测定了１９种杀虫剂对桑蚕的触杀、胃毒、熏毒性和残毒期评价了它们对桑蚕的安全性。     

Survival Analysis of 97 Cats with Nasal Lymphoma: A Multi-Institutional Retrospective Study (1986–2006)

Background: Feline nasal lymphoma (NLSA) is a condition for which no standard of care exists.
Hypothesis: There is no difference in survival times of cats with NLSA treated with single or multimodality therapy.
Animals: Records from 97 cats diagnosed with NLSA were examined.
Methods: The purpose of this retrospective study was to compare the survival times of cats with NLSA treated with radiation therapy (RT) alone, chemotherapy alone, or RT + chemotherapy and identify potential prognostic variables that affected survival. Cats were grouped according to therapy: RT + chemotherapy (n = 60), RT alone (n = 19), or chemotherapy alone (n = 18).
Results: Survival was calculated with 2 methods. The 1st survival analysis (method A) included all cats, but counted only deaths caused by progressive NLSA. The median survival time (MST), regardless of therapy modality, was 536 days. The 2nd survival analysis (method B) also included all cats and counted all deaths, regardless of cause, as events. The overall MST calculated for all deaths was 172 days. A negative independent prognostic variable identified was anemia ( P < .001), and positive independent prognostic variables were a complete response to therapy ( P < .001) and total radiation dose >32 Gy ( P = .03).
Conclusions and Clinical Importance: There were no significant differences in survival times among the 3 treatment groups but these results suggest that the addition of higher doses of RT to a cat's treatment protocol may control local disease and therefore influence survival.     

凝集试验检测鸡克雷伯氏杆菌病

本文建立了快速、简便、微量、高特异性的检测鸡克雷伯氏杆菌病玻片凝集试验。测得最适凝集原最适凝集原量为５０亿个菌／毫升，用来检测鸡克雷伯氏杆菌标准株和分离株的兔、抗血清效价分别为２５６－５１２（标准株）、５１２－１０２４（分离株）。临１６９分水岭分鸡粪样，８２５份鸡血清，阳性率分别为２８．４０％（４８／１６９）、２９．４５（２４３／８２５），而用细菌分离生化鉴定鸡粪阳性率为２９．５９（５０／１６９）     

Validation and Usefulness of the Sperm Quality Analyzer (SQA II-C) for Bull Semen Analysis

In this study, an upgrade version of the Sperm Quality Analyzer (SQA), the SQA-IIC was tested for the assessment of bull semen quality. In Expt 1, the device showed good repeatability of measurements within and between capillaries, as evidenced by the low coefficients of variation (CVs; < 13%) at concentrations between 35 and 705 x 10(6) spermatozoa/ml. In Expt 2, 10 semen concentrations (1-1000 x 10(6)/ml) were stored in HEPES TALP for 48 h at room temperature. A time-dependent decrease in sperm motility index (SMI) values was noticed. SMI values increased linearly with increasing sperm concentrations, but remained constant around 500, corresponding to a concentration of approximately 50 x 10(6)/ml. For sperm concentrations below 50 x 10(6)/ml, SMI values were highly correlated with concentration (p < 0.05) and with semen parameters, expressing the overall semen quality (p < 0.05; Expt 3). In Expt 4, a correlation of only 0.44 (p < 0.05) between SMI values of frozen-thawed semen samples of 35 bulls and the corrected 56-day non-return rate (56dNRRc) was found. Prediction of the 56dNRRc based on the SMI value of a semen sample was inaccurate. The present study indicates that the SQA-IIC is suitable for a rapid screening of bull semen diluted to a concentration of approximately 50 x 10(6)/ml. Furthermore, the device seems inappropriate for fertility prediction.