Suspected zinc-induced copper deficiency in growing kittens exposed to galvanised iron.

AIM: To investigate the possible causes of fading coat colour and an acquired hind-limb ataxia affecting sixteen 4 to 5-month old kittens in a closed feline colony during 1993 and 1994. METHODS: Records of kittens and litters born in the colony between 1991 and 1997 were analysed. The kittens had been kept from birth until approximately 5 months of age in plastic cages with galvanised iron bar doors. Histopathological sections from 4 of the worst affected ataxic kittens necropsied in 1993 were re-examined. In addition, 6 of the original 16 affected kittens that survived were re-examined as 4 to 5-year old adults, which were moderately ataxic; these cats were then humanely killed for necropsy. RESULTS: In the kittens, clinical signs included lordosis, dysmetria, ataxia of the hind-limbs and fading coat colour; histopathological lesions included Wallerian-type degeneration in the spinal cord, pons and medulla, and neuronal degeneration in the vestibular nuclei and ventral horns of the spinal cord. Analysis of colony data ruled out an inherited disease, and there was no evidence of dietary inadequacy or excess. Similar, though milder, clinical and histopathological changes were noted in the affected adults. CONCLUSIONS: Circumstantial evidence is consistent with a diagnosis of zinc-induced copper deficiency caused by the ingestion of zinc oxide from the galvanised iron bar doors. CLINICAL RELEVANCE: Because of the possibility of zinc-induced copper deficiency, galvanised iron should be avoided when designing and constructing cages for cats in veterinary clinics, pet shops and boarding facilities.

Relationship between plasma testosterone and urinary felinine in the growing kitten.

Felinine, a unique amino acid the functions of which are not well understood, is found in large quantities in male cat urine. Our study ran for 13 calendar months and involved taking monthly samples of blood and urine from 10 male and 10 female kittens starting at 6 months of age and measuring urinary felinine and plasma testosterone concentrations. Felinine was detectable at 6 months of age in all cats (range, mean +/- SEM, nmol/mL, 13.8-801.1, 432.3+/-112.2, males and 34.3-393.0, 140.4+/-45.0 females). In entire males, felinine showed a biphasic pattern, peaking (2550 nmol/mL) between 11-13 months of age toward the end of the attainment of puberty then falling to a low (1048 nmol/mL) at 15 months of age then climbing to a peak (3661 nmol/mL) at 17 months of age. Natural plasma testosterone levels in entire males showed a similar biphasic pattern peaking (6.8 pmol/mL) at 12 months of age, falling (1.3 pmol/mL) at 15 months, and finally rising again (12.6 pmol/ml) at 16 months of age. Castration of half the male cats induced a parallel fall in both testosterone and felinine that was reversed following testosterone supplementation. Urinary felinine levels in entire females rose slowly throughout the study and reached 795 nmol/mL at 18 months of age compared to the level of 365 nmol/mL reached by the spayed females: these levels were not significantly different when expressed as felinine/creatinine ratios. We could not detect testosterone in either entire or spayed females and so concluded that felinine was unrelated to testosterone in these groups. There was strong evidence that plasma testosterone levels are positively correlated with urinary felinine levels in male cats. The control of felinine might be sex-linked and may be part of a pheromonal signaling process of the male cat.

Urinary excretion of endogenous nitrogen metabolites in adult domestic cats using a protein-free diet and the regression technique.

The study was designed to determine urinary excretions of endogenous total, urea, ammonia and creatinine nitrogen in adult domestic cats. Endogenous urinary nitrogen metabolite excretions were determined by feeding adult cats a protein-free diet for 10 d or by regression to zero protein intake of the urinary nitrogen metabolite excretions of adult cats fed four levels of dietary protein. The mean (+/- SEM) endogenous total, urea and ammonia nitrogen excretions for the cats fed the protein-free diet were 360 (+/- 11.3), 243 (+/- 8.8) and 27.6 (+/- 1.06) mg x kg body weight(-0.75) x d(-1), respectively. Estimates of 316 (+/- 53.9), 232 (+/- 43.4) and 33.7 (+/- 5.68) mg x kg body weight(-0.75) x d(-1), respectively, were obtained using the regression technique. The differences in results between the two techniques were not statistically significant. Daily excretions of creatinine nitrogen were not significantly (P = 0.64) different between the protein-free and regression technique (mean +/- SEM, 14.4 +/- 0.49 and 15.9 +/- 1.05 mg/kg body weight(0.75), respectively). The endogenous urinary total and urea nitrogen excretion of adult domestic cats is higher than values for other mammals such as humans, dogs, rats and pigs.

Gut endogenous nitrogen and amino acid excretions in adult domestic cats fed a protein-free diet or an enzymatically hydrolyzed casein-based diet.

Ileal and fecal gut endogenous nitrogen and amino acid excretions in adult domestic cats were determined. Ileal digesta were collected (10 cm of terminal ileum) from the cats fed either a protein-free diet or an enzymatically hydrolyzed casein-based diet (free amino acids and peptides < 10,000 Da) for 1 wk. Chromic oxide was included in each diet as an indigestible marker. The relative contribution of the hindgut to total endogenous gut excretion was investigated in a separate study by feeding cats a protein-free diet with or without added antibiotics for 10 d. Endogenous ileal nitrogen and amino acid nitrogen excretions of (mean +/- SEM 2.4 +/- 0.27 and 1.9 +/- 0.13 mg/g food dry matter intake, respectively, were found for the cats fed the protein-free diet, whereas higher excretions of 3.6 +/- 0.73 (P = 0.12) and 3.6 +/- 0.76 (P = 0.03) mg/g food dry matter intake were obtained in cats fed the enzymatically hydrolyzed casein. Significantly (P < 0.05) higher endogenous ileal amino acid excretions, for the enzymatically hydrolyzed casein-fed cats compared with those fed the protein-free diet, were found for methionine, aspartic acid, serine, glutamic acid, proline, valine and isoleucine, with the differences in excretions of glycine, alanine, leucine and histidine being significant at the 6% level. Most of the endogenous fecal amino acid excretions were unaffected by the inclusion of the antibiotics in the protein-free diet, although bacterial numbers were significantly lower (69%). Antibiotics addition led to significantly higher fecal endogenous excretions of nitrogen, taurine, threonine, serine and histidine but significantly lower excretions of methionine and lysine. Cats, like other simple-stomached mammals, excrete higher amounts of endogenous amino acids at the terminal ileum when the diet contains peptides.

Felinine: a urinary amino acid of Felidae.

Felinine (2-amino-7-hydroxy-5,5-dimethyl-4-thiaheptanoic acid) has been identified in the urine of several members of the Felidae family including the cat (Felis catus). Rates of excretion of 95 mg/day have been recorded for entire male cats with entire female cats excreting around 19 mg/day. These high excretion rates in entire male cats may have a significant effect on the daily sulphur amino acid requirement. The isoamyl moiety of felinine seems to originate from the same isoprenoid pool as used for the synthesis of cholesterol in the cat. The sulphur in the felinine molecule appears to originate from cysteine, although some contradictory evidence exists. The site of synthesis and the method of transportation in the blood remain largely unknown. The biological significance of felinine to the animal is still a matter for speculation, but its function as a precursor to a pheromone seems likely. Recently, an accurate chemical assay for felinine has been developed that will allow investigation of felinine in different tissues and excretions and from a wider range of mammals.

Twenty-four hour felinine [corrected] excretion patterns in entire and castrated cats.

The purpose of this study was to determine the 24 h urinary excretion of a sulphur containing amino acid called felinine in entire and castrated cats of both sexes. Entire male cats excreted (mean +/- SEM) 122 +/- 23.6 mmol of felinine per kg bodyweight per day with castrated males, entire females and spayed females excreting 41 +/- 8.4, 36 +/- 7.3 and 20 +/- 3.8 mmol, respectively. There was an overall significant difference between groups in the amounts of felinine excreted in 24 h [F(3, 24) = 11.8, p < 0.0001] with there being significant differences between entire males and castrated males (p < 0.001) and castrated males and spayed females (p < 0.05). There was no difference in excretion between entire and spayed females. Urine volumes were not significantly different for the 24 h period. The differences in excretion levels were caused by different concentrations of felinine in the urine with entire male cats excreting (mean +/- SEM) 2.0 +/- 0.55 g of felinine per litre of urine. The data obtained in the present study support the concept that felinine, which has been found in Felidae species only, may be testosterone dependent. Felinine may be involved in territorial marking.

Serum free thyroxine levels in cats maintained on diets relatively high or low in iodine.

Excessive or deficient intake of iodine may play a role in the development of goitre and hyperthyroidism in cats. Previous investigations have shown that the serum free thyroxine level of cats is affected by brief administration of food high or low in iodine content. We have now measured serum free thyroxine levels in groups of cats fed relatively high or low iodine diets for much longer periods (5 months). In contrast to our earlier findings, the chronic ingestion of relatively high or low iodine diets did not lead to statistically significant differences in serum free thyroxine levels. The results of the present investigations show that the cat is able to maintain normal levels of thyroid hormone despite prolonged high or low iodine diets. It may be that the adaptive mechanisms that are called into play during chronic high or low iodine intake lead to thyroid disease, particularly in certain predisposed individuals.

Serum free thyroxine levels respond inversely to changes in levels of dietary iodine in the domestic cat.

Because of a perceived increase in the incidence of toxic multinodular goitres in cats in recent years, we investigated the iodine content of three varieties of commercial canned cat foods and studied the acute effects of 'ingestion of these preparations on urinary iodine excretion and serum free thyroxine levels in young, healthy cats. Ten castrated male cats were fed from a common source. The type of food was changed every 2 weeks. Urine and blood specimens were obtained weekly. Serum free thyroxine levels were determined and iodine concentrations in urine were assayed. The iodine content of the cats' food was also assayed. Food varieties of high, intermediate and low iodine content were fed for 2-week periods. There was a consistent, reciprocal relationship between the mean urinary iodine concentration and the mean serum free thyroxine level for each 2-week period. The difference in the mean serum free thyroxine concentrations for the high and low iodine intake periods was highly significant (p<0.01). When the serum free thyroxine level and the urinary iodine level for each cat at each collection throughout the 12-week study were analysed (66 paired results), a strong inverse correlation (r=0.59, p<0.01) was found. We concluded that the serum free thyroxine level in cats, as measured by a kit designed for human serum, is acutely responsive to changes in iodine intake.

Iodine content of commercially-prepared cat foods.

Twenty-eight varieties of commercially-available cat food (23 canned, 5 dried) were analysed for iodine. The iodine concentration varied from less than 0.37 micromol/kg to 41.8 pmol/kg, wet weight (less than 1.48 micromol/kg to 167 micromol/kg, dry weight). Excessive or insufficient iodine intake or wide swings in iodine intake over prolonged periods may contribute to thyroid disorders in cats.

Feline struvite crystalluria: a comparison of some commercially available dry cat diets in New Zealand.

Thirteen adult cats were each fed consecutively five test diets: firstly a complete canned diet, and then four commercially available dry cat diets (New Formula Biscats, Cat Chow, Go-Cat and Whiskettes) for 7,13, 5, 4 and 4 weeks respectively. Urine samples were tested for pH and presence of struvite crystals. Body weight was recorded weekly. Urine pH was ranked from acid to alkaline as follows: Whiskettes (pH 5.8), canned diet (pH 5.9), Biscats (pH 6.1), Cat Chow (pH 6.45) and Go-Cat (pH 6.5). The pH of urine when cats were fed Whiskettes, Biscats and canned diet was not significantly different. Biscats produced a urine pH significantly higher than Whiskettes but, in each case, these three diets produced a significantly lower urine pH than Cat Chow and Go-Cat. Struvite crystals occurred spontaneously in 9% of urine samples from cats when fed Whiskettes and Biscats, 71% when fed Cat Chow and 38% when fed Go-Cat: spontaneous struvite was not seen in any cat fed CX9. Haematuria was seen in 2.9% (2/69) urine samples from cats fed Cat Chow. One tom cat had a blocked urethra after 24 days on Go-Cat. No abnormalities were observed in any cat when fed canned diet, Biscats or Whiskettes. It was concluded that of the four dry cat diets tested only Biscats and Whiskettes produce urine with an acceptably low rate of spontaneous struvite crystalluria.

Postnatal influence of diethylstilbestrol on the differentiation of the sexually dimorphic nucleus in the rat is as effective as perinatal treatment.

The volume of the sexually dimorphic nucleus of the preoptic area (SDN-POA) of the male rat brain is larger than that of the female. In the female rat, treatment with diethylstilbestrol (DES), either perinatally (from day 16 of gestation to postnatal day 10), or postnatally (birth to day 10) was equally effective in increasing the volume of SDN-POA compared to controls. Prenatal treatment (day 16 of gestation to birth) with DES also increased the volume of the SDN-POA but this increase was significantly smaller than that achieved with the other treatments. These results confirm the effectiveness of DES in increasing the volume of the SDN-POA in the female rat brain, and prove that the differentiating SDN-POA is very receptive to hormone influences in the early postnatal period.

Feline struvite urolithiasis: fasting reduced the effectiveness of a urinary acidifier (ammonium chloride) and increased the intake of a low magnesium diet.

In three separate experiments nine male cats were fed either a canned complete diet or a commercially available dry pelleted diet or the same dry diet containing 1.6 per cent ammonium chloride for seven days and then fasted for 20 hours. Then ad libitum feeding was continued and urine samples were taken at four-hour intervals for 12 hours and a final sample 12 hours later. Urine pH and the presence of struvite crystals in urine sediment were evaluated. The food and water intake of four of the nine cats was measured at the time of urine collection. After the fast, urine pH was raised, even after feeding the dry diet supplemented with ammonium chloride. A post prandial rise in urine pH was also seen on all three diets. After feeding the dry diet the postprandial peak pH was 7.97 and struvite occurred spontaneously. Urine pH after feeding the dry diet supplemented with ammonium chloride peaked at 7.75 then fell to 6.1 12 hours after the start of feeding. Struvite occurred spontaneously at all times until the pH reached 6.1 but when the pH of urine was raised to 7.0 the struvite crystallised. Urine pH on the canned complete diet peaked at 6.8 then fell to 5.8; struvite did not occur spontaneously but when urine pH was raised to 7.0 struvite crystallised except at the eighth and 12th hour sampling. These data show that fasting initiates a post prandial rise in urine pH and struvite crystalluria even when a normally effective urinary acidifier is used.(ABSTRACT TRUNCATED AT 250 WORDS)

Feline struvite urolithiasis: factors affecting urine pH may be more important than magnesium levels in food.

Eighteen male cats were fed either a canned complete diet or a commercially available dry pelleted diet or the same dry diet containing 1.6 per cent ammonium chloride. The daily food and water intake of four of these cats was measured. Urine samples were taken at random and the pH and the presence of struvite crystals in their sediment estimated. In some samples in which the pH was less than 7.0, and struvite crystals were absent, the pH was increased to 7.0 and any sediment was examined for struvite. No spontaneous struvite was seen in urine samples (pH 5.8) from cats fed the canned complete diet but when its pH was raised to 7.0, 46 per cent of these samples showed struvite. Cats fed the dry pelleted diet had urine of higher pH (7.55) and 78 per cent of the samples contained struvite crystals. Cats fed this diet supplemented with ammonium chloride had a urine pH of 5.97 and only 9 per cent contained struvite crystals but when the pH was adjusted to 7.0 all the samples showed struvite crystals. Energy intake was similar on all three diets but the intake of dry matter was greater on the dry diets. Liquid water intake was greater on the dry diets but total water intake was greatest on the canned complete diet. The intake of magnesium, calcium and phosphate was greater on the dry diets. It is concluded that urine pH is a more important controller of struvite precipitation than mineral intake.

Early prenatal treatment of ewes with testosterone completely masculinises external genitalia of female offspring but has no effects on early body weight changes.

Treatment of pregnant ewes from day 20 of gestation with 100 mg implants of crystalline testosterone did not cause masculinisation of genitalia or affect growth rates of female lambs. Prenatal treatment from day 20 of gestation with testosterone propionate (1.2 g in divided doses for four weeks) or testosterone cypionate (600 mg in 3 doses over three weeks) completely masculinised the external genitalia of female lambs producing a ventral penis and scrotum with no external vaginal opening: ovaries and uteri were normal. Single male lambs were significantly heavier than female lambs during a 16 week measurement period, but masculinised female lambs were not different from controls. In the twin lamb groups, there were no differences between male and female or treated and control lambs. Body weights of castrated male lambs born as twins were not different from entire controls. It was concluded that testosterone treatment of developing lambs from 20 to 65 days of gestation produces complete masculinisation of external genitalia of female lambs but does not affect body weight during the first 16 weeks of age.

Plasma testosterone levels in adult and neonatal female rats bearing testosterone propionate-filled silicone elastomer capsules for varying periods of time.

Testosterone propionate (TP) was administered, by means of subcutaneous implanted silicone elastomer capsules, into adult and neonatal (aged 3 days) female rats. In the adult rats a dose-dependent increase in plasma testosterone was measured for capsules of three different sizes (5, 10 and 20 mm crystal length). Testosterone levels reached a peak 4-8 h after insertion (5 mm, 24.6 +/- 1.4 (S.E.M.) nmol/l; 10 mm, 34.0 +/- 3.8; 20 mm, 44.4 +/- 3.1) and returned to control levels within 4 h after removal: the calculated half-life of testosterone was 1 h for all sizes of capsule. In neonates, a capsule of 2.5 mm crystal length was removed after 4 h subcutaneous implantation (at day 3 of age) and produced peak testosterone levels of 126.2 +/- 11.8 nmol/l: the calculated half-life was 8.6 h which compared with a half-life of 48 h after a subcutaneous injection of 312 mumol TP (in 0.05 ml arachis oil) which produced peak levels of testosterone in 4-8 h of 84.6 +/- 11.8 nmol/l. Chronic implants of TP-filled capsules (2.5 mm crystal length) at 3 days of age and left in situ for 15 weeks gave a half-life of 69 h. Removable silicone elastomer capsules were found to be a versatile vehicle for the administration of TP to rats of all ages where precise hormone treatment for a known period or prolonged administration is required. The duration and magnitude of plasma hormone levels should be established by assay in an in-vivo situation.

Some effects of ovariectomy and estrogen replacement on body composition in the rat.

Sprague-Dawley rats were ovariectomized (OvX) at 3 ages, day 2 (D2), week 4 (W4) and week 7 (W7); a group of OvX W7 rats were treated daily with estrogen (OB;2 micrograms for 2 or 5 weeks from 10 weeks of age). Rats were slaughtered at 4 ages, weeks 7, 9, 12 and 15, for the chemical analysis of carcass and skin. Chemical compositions were analysed as % wet weight and as component weights by two-way analysis of variance. Component weights were also analysed by allometry, regressing against nose-anal length. Ovariectomy increased overall body weight without causing obesity. The weight gain of the OvX rat was mainly a true growth response but OvX affected body proportions so that at a given body length the OvX rat had a larger skin and carcass than controls. Ovariectomy at the earliest age (D2) produced the smallest response in body weight and body length but produced the greatest fat redistribution towards the skin and away from the carcass; there was no net change in whole body fat levels following OvX. Long-term daily OB treatment increased fat reserves but slowed the growth of other body components, including the axial skeleton. Whereas OvX redistributed components between skin and carcass, OB treatment reversed this process.

Use of perfluoroalkanoic acids as volatile ion pairing reagents in preparative HPLC.

This report describes the successful use of the ether soluble, ion pairing reagents perfluoropropoinic and perfluorobutyric acid in the preparative and analytical reserved phase HPLC of underivatised peptides. The preparative separation of a 1-g sample of Pyr-His-Gly, the proposed anorexigenic peptide, is described on C18-silica which was packed in a flexible-walled cartridge and subjected to radial compression. The mobile phase consisted of an aqueous solution of perfluorobutyric acid (5 mM) and a flow rate of 100 ml/min was used. The purified peptide was simply isolated by neutralizing and freeze-drying the corresponding peak and then extracting the excess ion pairing reagent with ether. The product was then shown to be homogeneous by analytical HPLC and amino acid analysis. The tripeptide failed to show any effect on food intake, water intake or body weight in female rats. Similarly no effect was noted on the reproductive cycles of the rat.