Vaginal and Cervical Anatomic Modifications During the Oestrus Cycle in Relation to Transcervical Catheterization in the Domestic Cat

In a previous study we observed that it is possible to reach the cervix in all queens with a 1 mm diameter probe only. So, we developed both a new technique and a catheter (1 mm diameter) to allow transcervical insemination [Zambelli and Castagnetti 2001]. The aims of this study were to investigate vaginal and cervical anatomic modifications during the various stages of the oestrus cycle and to test the previously described technique of transcervical catheterization during the various stages of the oestrus cycle. In experiment 1, silicon impression moulds were obtained from the reproductive tracts of 21 queens' cadavers and vaginal and cervical measures were taken. The results showed that there are some significant anatomic modifications during the various stages of the oestrus cycle in vaginal and cervical anatomy, principally related to the dorsal medial fold increase induced by the follicular phase. In experiment 2, transcervical catheterization was attempted in 95 queens at various stages of oestrus cycle both during reproductive and non-reproductive season. After catheterization, methylene blue solution was injected through the cervical catheter. Successful catheterization was assessed during surgery, when colour was observed in the uterine horns. It was possible to perform transcervical catheterization during non-reproductive season in 16 of 20 anoestrus queens and in 12 of 15 induced oestrus queens; during reproductive season in nine of 21 interoestrus queens, in eight of 13 dioestrus/pregnancy queens, in four of 18 oestrus queens and in seven of eight queens in first oestrus during lactation.     

Ultrastructural Characteristics of Non-matured and In Vitro Matured Oocytes Collected from Pre-pubertal and Adult Domestic Cat Ovaries

The present study describes the ultrastructural characteristics of cat oocytes before maturation and after 12- and 24-h in vitro maturation (IVM). Oocytes were recovered from pre-pubertal and adult queen ovaries after ovariohysterectomy and a proportion were stored in glutaraldehyde at 4°C until examination by transmission electronic microscopy (TEM). Those selected for maturation were cultured before TEM in DMEM for 12 and 24 h at 38°C in a humidified environment of 5% O₂, 5% CO₂ and 90% N₂. Specimens were divided into six groups: non-matured oocytes from pre-pubertal queens (PP0), non-matured oocytes from adult queens (A0), 12-h in vitro matured oocytes from pre-pubertal queens (PP12), 12-h in vitro matured oocytes from adult queens (A12), 24-h in vitro matured oocytes from pre-pubertal queens (PP24) and 24-h in vitro matured oocytes from adult queens (A24). Across the treatment groups, it was possible to observe differences in the thickness of the perivitelline space, the penetration of cumulus cell projections forming a junctional complex, distribution and density of small vesicles, lipid droplets, microvilli, mitochondria and cortical granules and variable degrees of development of Golgi complexes. These findings demonstrated that ultrastructural analysis of oocytes matured in vitro is a valuable tool to evaluate oocyte cytoplasmic maturation and that this IVM protocol was efficient in inducing gradual morphological changes necessary for cytoplasmic maturation of pre-pubertal and adult cat oocytes.     

Adrenocortical function in the domestic cat during treatment with levonorgestrel

Levonorgestrel was administered via a subcutaneous, slow-release silastic implant to 10 queens. Five other queens served as controls. Their adrenocortical function was assessed by the adrenocorticotrophic hormone (ACTH) stimulation test before and after one, two, six and 12 months of treatment. In addition, the gross anatomy and histology of the adrenal gland were examined post mortem in six of the treated cats. In both the control and treated queens the plasma cortisol concentrations (pre and post ACTH) were significantly different (P<0·05) at different times. However, there were no significant differences between the plasma cortisol concentrations (pre and post ACTH) of the treated and control queens. No gross or microscopical abnormalities were visible in the adrenal glands of the treated queens.     

饲粮粗蛋白质水平对中华蜜蜂育王质量的影响

本试验旨在研究饲粮粗蛋白质水平对中华蜜蜂蜂王育王的影响。选取基本情况相近的中华蜜蜂9群,随机分成3组,每组3群,在每群蜂巢门口装置脱粉器,阻止花粉进入,使其处于缺粉状态。Ⅰ组、Ⅱ组、Ⅲ组蜂群分别饲喂粗蛋白质水平为15%、25%、35%的试验饲粮,45 d后控制蜂王产卵,并进行人工育王,待蜂王出房后,测其初生重、胸宽,并检测其卵巢卵黄蛋白原基因(vitellogenin,Vg)、转铁蛋白基因(transferrin,Trf)的相对表达量。结果表明:蜂王的初生重随饲粮粗蛋白质水平的增加而升高,Ⅲ组和Ⅱ组培育蜂王的初生重显著高于Ⅰ组(P0.05),且Ⅲ组也显著高于Ⅱ组(P0.05);蜂王的胸宽随饲粮粗蛋白质水平的增加而升高,但3个试验组之间差异不显著(P0.05)。Ⅲ组和Ⅱ组培育蜂王卵巢的Vg相对表达量显著高于Ⅰ组(P0.05),但Ⅲ组与Ⅱ组之间差异不显著(P0.05);培育蜂王卵巢的Trf相对表达量3个试验组间均有显著差异(P0.05),相对表达量最高的是Ⅲ组,最低的是Ⅰ组。由此得出,饲粮粗蛋白质水平对中华蜜蜂育王质量存在影响,高蛋白质饲粮可以增加中华蜜蜂蜂王的初生重,促进蜂王卵巢Vg和Trf的表达。育王时,中华蜜蜂饲粮粗蛋白质水平不应低于25%。     

Infertility in the cycling queen: seven cases

In this study we examined seven queens with normal oestrous cycles and a history of infertility after normal matings. We performed clinical examination, vaginal cytology, evaluation of oestradiol, progesterone and total T4 levels, vaginal bacterial culture, ultrasonography, and serum analyses for detection of antibodies against chlamydia and feline leukaemia virus (FeLV) antigenemia. Ovariohysterectomy (OHE) was recommended for 1/7 queens because of pathological uterine changes detected at ultrasonography and clinical examination. Four out of seven queens were treated with antibiotics and two of these had more litters. One of the queens that were treated was not mated again and one was mated without conceiving and was at a later OHE found to have degenerative uterine changes. No treatment was given and no diagnosis could be established in 2/7 queens. Both of them were later ovariohysterectomised and one showed degenerative uterine changes while the uterus of the other queen could not be obtained for follow-up. In summary, 4/7 cats were diagnosed with uterine pathology and no definitive diagnosis could be established for 3/7 queens.     

Effects of passive transfer of immunity on results of diagnostic tests for antibodies against feline immunodeficiency virus in kittens born to vaccinated queens

OBJECTIVE: To determine whether passive transfer of immunity affects results of diagnostic tests for antibodies against FIV in kittens born to vaccinated queens. DESIGN: Experimental trial. ANIMALS: 12 specific-pathogen-free queens and their 55 kittens. PROCEDURE: Queens were vaccinated with a whole-virus FIV vaccine prior to breeding. Serum was obtained from the queens on the day of parturition and from the kittens on days 2 and 7, then weekly until results of tests for antibodies against FIV were negative for 2 consecutive weeks. Milk was collected from the queens daily for the first week and then weekly. Serum and milk were tested for antibodies against FIV with 2 commercial assays. RESULTS: Antibodies against FIV were detected in serum obtained from the queens on the day of parturition and in the milk throughout lactation. All kittens tested positive for antibodies against FIV at 2 days of age. At 8 weeks of age, 30 (55%) kittens tested positive with 1 of the commercial assays, and 35 (64%) tested positive with the other. All kittens tested negative for antibodies against FIV by 12 weeks of age. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that kittens readily absorb antibodies against FIV in colostrum from vaccinated queens and that these antibodies may interfere with results of commercially available tests for FIV infection past the age of weaning. Currently licensed diagnostic tests for FIV infection are unable to distinguish among kittens with antibodies against FIV as a result of infection, passive transfer from infected queens, and passive transfer from vaccinated queens.     

Feline embryo transfer during the non-breeding season

To obtain normal kits by embryo treansfer (ET) during the non-breeding season, maintenance of pregnancy was carried out by administration of sustained action progesterone (P4) in queens. Embryos were recovered six days after mating from five donor queens in which ovulation was induced by administration of eCG and hCG. The number of embryos recovered ranged from 24 to 53 (mean: 37.2 +/- 6.4) per animal and most embryos were compacted morulae. The yield of embryos was 49.0-93.3% (mean: 73.8 +/- 9.6%). As for recipients, porcine pituitary gland preparation and hCG were administered to 19 queens and estrus and ovulation were induced in 18 queens (94.7%). These queens underwent intrauterine ET of five compacted morulae and 17 cats (94.4%) were impregnated. The number of implantations was 2-5 (mean: 3.7 +/- 0.3). Among these impregnated queens, 15 cats received P4 adminstration starting on day 24 of gestation and 1-5 newborns (mean: 3.4 +/- 0.3) were obtained by normal delivery or caesarean section on day 64-69 of gestation. However, two animals that were not treated with P4 underwent spontaneous abortion about the mid gestational period. Therefore, it is possible to obtain normal kits from queens in the non-breeding season by ET with maintenance of pregnancy by P4 administration.     

Immunoglobulin concentrations in feline colostrum and milk, and the requirement of colostrum for passive transfer of immunity to neonatal kittens

The purpose of this study was to clarify whether cats have a colostral and milk phase of lactation differentiated by concentrations of immunoglobulins, and whether colostrum ingestion by newborn kittens is essential for optimal transfer of passive immunity. Milk from specific pathogen-free queens was analyzed for IgG and IgA concentrations from parturition through 6 weeks of lactation. Serum IgG and IgA concentrations from birth through 8 weeks of age were determined for colostrum-fed kittens, colostrum-deprived kittens that were fed a milk replacer, and colostrum-deprived kittens that were fostered onto queens in the milk phase of lactation. The total IgG and IgA concentrations in milk were significantly higher on the day of parturition than on day 7 of lactation, indicating cats do have a colostral phase of lactation. The predominant immunoglobulin in both colostrum and milk was IgG. The serum IgG concentrations in colostrum-deprived kittens fostered on queens in the milk phase of lactation were similar to colostrum-deprived kittens fed a milk replacer, and the concentrations were significantly lower than in colostrum-fed kittens for the first 4 weeks of life. The serum IgA concentrations in both colostrum-deprived groups were significantly lower than colostrum-fed kittens on day 2 after parturition, but were similar thereafter. Colostrum-deprived kittens fostered onto queens in the milk phase of lactation had failure of passive transfer of maternal antibodies. Protective concentrations of immunoglobulins can be restored in kittens with failure of passive transfer of immunity by parenteral administration of adult cat serum, but not by fostering on queens in mid-lactation.     

Long-term prevention of estrus in the bitch and queen using chlormadinone acetate

Estrus was prevented with weekly oral administration of 2 mg chlormadinone acetate for 2.0 to 9.8 y in bitches and queens. Abnormalities, including mammary or uterine disorders, or both, were noted in 7 out of 14 bitches and 9 out of 24 queens during this long-term treatment.     

Chronic ingestion of high concentrations of cholecalciferol in cats

OBJECTIVE: To determine whether ingestion of 63 times the recommended amount of vitamin D3 (cholecalciferol) results in renal calcification or damage in cats. ANIMALS: 20 four-month-old kittens, 17 queens, and 20 kittens born to these queens. PROCEDURE: 4-month-old kittens and queens were given a purified diet with 846 microg of cholecalciferol/kg of diet (high vitamin D3 diet) or 118 microg of cholecalciferol/kg of diet (control diet) for 18 months. Kittens born to queens were weaned onto the same diet given to dams. RESULTS: There were no apparent adverse effects of the high vitamin D3 diet. Plasma cholecalciferol and 25-hydroxycholecalciferol (25-OHD3) concentrations of queens and 4-month-old kittens given the high vitamin D3 diet significantly increased with time. At 6 months, plasma cholecalciferol concentrations in these kittens and queens were 140.0+/-7.3 nmol/L and 423.6+/-26.6 nmol/L, respectively (10 times initial values). Corresponding 25-OHD3 concentration in queens was 587.5+/-59.4 nmol/L (2.5-fold increase over initial values). At 3 months of age, kittens born to queens given the high vitamin D3 diet had an increase in serum BUN and calcium concentrations and a decrease in RBC and serum total protein, albumin, and hemoglobin concentrations. By 18 months, these kittens had an increase in plasma cholecalciferol (276.0+/-22.2 nmol/L) and 25-OHD3 (1,071.9+/-115.3 nmol/L) concentrations. However, all indices of renal function and the appearance of renal tissue on histologic evaluation were normal. CONCLUSIONS AND CLINICAL RELEVANCE: These results indicate that cats are resistant to cholecalciferol toxicosis when the diet is otherwise complete and balanced.     

The use of bipolar electrosurgical forceps for haemostasis in open surgical ovariectomy of bitches and queens and castration of dogs

Objectives

To describe a technique using bipolar electrosurgical forceps for haemostasis during open ovariectomy of bitches and queens and for castration of dogs and to determine whether these forceps reduced surgical time compared to ligation with suture.

Materials and Methods

Bipolar electrosurgical forceps were used for haemostasis in 3744 open surgeries including ovariectomies in bitches (n=1406) and queens (n=859) and castrations in dogs (n=1335). The forceps were also used to assist with ovariohysterectomy in bitches (n=89) and queens (n=55). The effect of bipolar electrocoagulation and other likely factors influencing surgical time was examined using linear regression analysis of 367 surgeries.

Results

The use of the forceps reduced surgical time by 9·7 ±2·8 minutes in bitches and by 3·0 ±1·5 minutes in queens. The complications encountered initially were small superficial skin burns requiring topical treatment and one case of haemorrhage associated with surgical technique.

Clinical Significance

Bipolar electrosurgical forceps reduce surgical time in ovariectomies, and complications are rare. This technique has promise for widespread application in veterinary practice to assist with haemostasis in ovariectomy of bitches and queens.     

Cats during gestation and lactation fed with canned food ad libitum: energy and protein intake, development of body weight and body composition

The NRC recommendations for cats for energy and protein supply during gestation and lactation are based on limited data. This study aimed to answer the question: Can the energy requirement be met with canned food or is the volume restrictive? Therefore, balance trials were conducted in 10 queens before mating, during the 4th and 7th week of gestation and during the 2nd and 6th week of lactation. The cats were fed with canned food ad libitum. Additionally, the body composition of the queens was measured by dual‐energy X‐ray absorptiometry (Dexa) before mating, after parturition and after weaning. Eight of 10 cats presented increased body fat content and lean body mass during gestation. The weight loss during lactation led to a loss of lean body mass, but only six cats lost body fat of widely differing amounts. It was evident that the queens’ dry matter intake was consistent with that of queens fed ad libitum with dry food. The cats lost lean body mass during lactation and had negative protein balances in the 2nd week of lactation. This seems to be physiological in early lactation. Nevertheless, the protein recommendations for lactation seem to be too low.     

ULTRASOUND PREDICTION OF PARTURITION IN QUEENS

Eight pregnancies in five queens were followed with serial ultrasound examinations. Measurements of head and body diameter were made throughout pregnancy. These measurements were used to produce individual growth curves for each pregnancy and average curves for both head and body diameter. Formulas based on the slope of the growth curves were derived. These formulas were then used to predict parturition dates based on single measurements of fetal head and/or body diameter in eight additional queens. Predicted date of parturition was then compared to actual queening dates. Ultrasound prediction of parturition date was accurate to within two days in seven of eight queens.     

Prevention of estrus in the queen with chlormadinone acetate administered orally.

The possibility of estrus prevention in the queen by the oral administration of chlormadinone acetate was examined. The animals used were 29 mature and 15 immature queens. For 16 mature females, 4-12.5 mg was given daily by mouth for 7 days every 3 months. Ten of the 16 queens given this treatment came into estrus within 4 months of the first treatment. For 28 females including the immature, 2-12.5 mg was given once a week throughout the experiment. This treatment prevented estrous activity for at least 1 year. In the queens in this study, the side effects were not observed excepting an increase in body weight during treatment. Our results showed that oral administration of this drug weekly is safe and reliable for long-range prevention of estrus in queens.     

A survey of sexual behaviour and reproduction of female cats

Details of reproductive activity among 168 breeding queens were obtained from a survey of specific information supplied by cat breeders. Puberty was reached, on average, at 9–10 months of age. The gestation period was calculated to average 65 days. Almost half the queens in the survey showed cyclic occurrence of oestrus throughout the year. Cycles were usually 21 days long. When a non-breeding season was present it generally occurred between September and the end of January, its average duration being 4 months. The data obtained from the survey are examined and discussed, and are considered to illuminate usefully a scantily reported topic.     

Investigation of Cervical Patency and Uterine Appearance in Domestic Cats by Fluoroscopy and Scintigraphy

The cervical patency of six domestic female cats was monitored under sedation by infusion of contrast medium (Omnipaque) into the cranial vagina during early oestrus, mid‐oestrus, late oestrus and interoestrus or a radiopharmaceutical (^99mTc‐HSA) during mid‐ and interoestrus in a non‐ovulatory oestrous cycle. The transport of the contrast medium or the radiopharmaceutical through the cervix and within the uterine horns was observed under fluoroscopy and with the aid of scintigraphy. In three of the queens, transcervical transport of contrast medium was demonstrated in all stages of oestrus, in one queen during mid‐oestrus, late oestrus and 1 day after oestrus, and in two queens only during late oestrus. The relations between the cervical patency to the contrast medium and the oestrous behaviour, cornification of the vaginal cells and the serum oestradiol‐17β concentration were evaluated, and a relationship was found between the cervical patency and the degree of vaginal cornification. Transcervical transport of the radiopharmaceutical was observed in three queens during mid‐oestrus. When the cervix was open, hysterography under a fluoroscope and hysteroscintigraphy were performed. The fluoroscopic and scintigraphic recordings revealed the patterns of the uterine contractions during oestrus in both ascending and descending directions, and the movement of the uterine contents back and forth between the uterine horns. The hysterograms were classified according to the shape of the uterine horns and the appearance of the endometrial lining. Spiral‐shaped uterine horns with a smooth inner contour were observed in two queens, and a corkscrew appearance with irregular filling defects in the uterine lumen was shown in two queens that had developed subclinical cystic endometrial hyperplasia. These findings demonstrated that fluids or particles deposited in the cranial vagina of the cat can be transported into the uterus during some stages of the oestrous cycle. The fluoroscopic and scintigraphic techniques developed in this study may be further modified to permit more detailed studies of uterine contractile patterns and sperm transport in the feline female reproductive tract. Hysterography proved useful to diagnose uterine disease. The information on cervical patency is of value also for the development of techniques for artificial insemination in this species, and should be studied also in the ovulatory cycle.     

Feline neonatal isoerythrolysis in two litters

The clinical, pathological and serological findings for feline neonatal isoerythrolysis (FNI) in two litters are described. In each litter one of four kittens died shortly after birth, two showed varying degrees of depression, jaundice and haemoglobinuria and one remained healthy (clinically unaffected). Three of the kittens with jaundice died and the necropsy findings included variable degrees of erythrophagocytosis in the spleen and liver and many casts containing haemoglobin in the tubules and collecting ducts of the kidney. Blood typing and haemocompatibility testing were performed on the surviving kittens, queens and sires. Both queens and the healthy kittens were Type B while both sires and the surviving affected kitten were Type A. In both litters the queens sera haemolysed the sires erythrocytes but did not affect the erythrocytes of the healthy kittens. The erythrocytes of the surviving affected kitten were haemolysed by the serum of its queen. The pathogenesis and treatment of FNI are discussed.     

Comparison of steroid receptor expression in normal, dysplastic, and neoplastic canine and feline mammary tissues

Steroid receptor expression was assessed by immunohistochemistry in neoplastic, hyperplastic/dysplastic, and normal mammary tissue samples removed from 68 queens and 47 bitches, using monoclonal antibodies against human oestrogen-alpha (ER) and progesterone receptors (PR). Mammary lesions were classified according to World Health Organization (WHO) criteria, and all animals with invasive carcinomas were clinically followed for 2 years. Stromal and/or lymphatic invasion and histological grading were also recorded. In both species, ER expression was significantly higher in healthy tissues, hyperplastic/dysplastic lesions, and benign tumours than in carcinomas. The loss of ER expression was more marked in feline than in canine carcinomas. In queens, PR expression increased in dysplastic lesions and "in situ" carcinomas and decreased in invasive carcinomas, even if parts of these tumours were still PR-positive. In bitches no significant variation in PR expression was observed between normal tissue, dysplasias, and benign neoplasms, but was significantly lower in carcinomas. In both species ER and PR expression in invasive carcinomas did not correlate either with histological parameters or overall survival time. This study demonstrates several differences in steroid hormone dependency between the two species. The percentage of PR-positive feline carcinomas suggests a possible role of progesterone in promoting early tumour cell growth in queens. The low percentage of ER-positive invasive carcinomas further demonstrated the aggressive phenotype and behaviour of feline mammary tumours.     

意大利蜂工蜂,蜂王营养成分分析及其变化规律的研究

意大利工蜂和蜂王都大约在第９日龄（自卵产下开始计时）体重达到最大，此时蜂王平均体重约为工蜂平均体重的１８６７倍。蜂王在第７日到第８日龄期间生长特别快。蜂王的干物质、灰分含量比相应的工蜂要高，而总糖含量比相应的工蜂要低。工蜂、蜂王体内蛋白质的含量有波动现象，粗脂肪在工蜂、蜂王体内的含量及变化反映了蜂王的营养水平比工蜂高。意大利工蜂与蜂王的体重及干物质、灰分、总糖、粗脂肪、粗蛋白含量随发育时间变化规律都分别具有一定的相似性