Melanocortin-4 receptor and proopiomelanocortin: Candidate genes for obesity in domestic shorthair cats.

Obesity is an escalating global health problem affecting both humans and companion animals. In cats it is associated with increased mortality and multiple diseases, including diabetes mellitus. Two genes coding for proteins known to play a critical role in energy homeostasis across species are the proopiomelanocortin (POMC) gene and the melanocortin-4 receptor (MC4R) gene. A missense variant in the coding sequence of the feline MC4R (MC4R:c.92C>T) has been reported to be associated with diabetes and overweight in domestic shorthair cats, and while variants in the POMC gene are known to cause obesity in humans and dogs, variants in POMC and their association with feline obesity and diabetes mellitus have not been investigated to date. The current study aimed to assess the association between the previously described MC4R variant and body condition score (BCS), as well as body fat content (%BF) in 89 non-diabetic domestic shorthair cats. Furthermore, we investigated the feline POMC gene as a potential candidate gene for obesity. Our results indicate that the MC4R:c.92C>T polymorphism is not associated with BCS or %BF in non-diabetic domestic shorthair cats. The mutation analysis of all POMC exons identified two missense variants, with a variant in exon 1 (c.28G>C; p.G10R) predicted to be damaging. The variant was subsequently assessed in all 89 cats, and cats heterozygous for the variant had a significantly increased body condition score (p = 0.03) compared with cats homozygous for the wild-type allele. Results from our study provide additional evidence that the previously described variant in MC4R is not associated with obesity in domestic shorthair cats. More importantly, we have identified a novel variant in the POMC gene, which might play a role in increased body condition score and body fat content in domestic shorthair cats.

Evaluation of an in vitro system to simulate equine foregut digestion and the influence of acidity on protein and fructan degradation in the horse's stomach.

The aim of this study was to improve an in vitro system in order to gather optimized information on the digestion of different forages in the horse's upper gastrointestinal tract. Therefore, foregut digestion of several forages was simulated in vitro (Part 1). The effect of different pH values on in vitro fructan degradation of two selected grasses (Part 2) was tested subsequently. Part 1: We hypothesized that our system produces representative results simulating digestive processes in the upper alimentary tract, but neglects microbial fermentation. In vitro digestion of six forages (grass mixture for horses, grass mixture for cows (GMC), tall fescue, English perennial ryegrass (ER), white clover, lucerne) was performed in two phases with pepsin and pancreatin. The results are consistent with current data from in vivo studies, including a degradation of crude protein and monosaccharides as well as a relative increase in fibres. Interestingly, a loss of fructan was measured in two feedstuffs (ER/GMC: 4.1/4.4% DM fructan before and 0.59/0.00% DM after simulated foregut digestion). Part 2: As fructans are thought not to be fragmented by digestive enzymes, another hypothesis was developed: acidic hydrolysis leads to a degradation of fructans. To evaluate the influence of gastric pH on the digestion of fructan and protein, different pH values (2, 3 and 4) were adjusted in a second series of in vitro foregut digestion trials with ER and GMC. As expected, the highest degradation of protein was seen at the lowest pH (protein in ER/GMC at pH 2: 6.11/8.28% DM and at pH 4: 7.73/10.64% DM), whereas fructan degradation was highest at pH 4 (fructan in ER/GMC at pH 2: 1.63/1.95% DM and at pH 4: 1.31/0.91% DM). We presume that not only acidic hydrolysis but also plant enzymes cause the loss of fructans in an acidic environment.

[Investigation of metabolic parame- ters in high yielding dairy cows in pasture based production systems]. / Stoffwechseluntersuchungen bei Hochleistungskühen in Vollweidehaltung.

In the present study differences in metabolism between New Zealand Holstein-Friesian (NZHF) and Brown Swiss (CH-BV) or Swiss Holstein-Friesian (CH-HF) were investigated in a grassland based milk production system in Switzerland. Therefore 14 pairs of CH-BV/NZHF and 11 pairs of CH-HF/NZHF were available. The parameters glucose, insulin, non-esterified fatty acids (NEFA), ß-hydroxybutyrate (ß-HB), urea and cholesterol were analysed at the times 5-3 weeks before the calculated partus and 2, 3, 5, 7, 10 and 18-22 weeks post partum. Only ß-HB showed significantly higher concentrations (P = 0.0059) for both Swiss breeds compared to the NZ-HF. Regarding all other physiological parameters during early lactation New Zealand Holstein-Friesians were not different from Swiss breeds.

Feeding practice and influence on selected blood parameters in show jumping horses competing in Switzerland.

The aim of this study was to compare the nutritional management of show jumping horses in practice with recommendations from the literature. Additionally, the effects of these feeding practices on several blood metabolic parameters before and after exercise were studied. Blood samples were collected in the field from 27 different horses at 71 trials on the level M1 to S2 show jumping competitions in Switzerland and questionnaires on feeding practice of the studied horses were evaluated. The questionnaires revealed that during training and on tournament days horses received on average 3.1 kg of concentrate per day (min. 2.0 kg, max. 6.6 kg) divided into two to three meals. The horses were fed on average 6.9 kg of roughage per day (min. 4.0 kg, max. 13.0 kg). Additionally, it was observed that the horses received the last meal on average 6 h 10 min (min. 1 h 50 min, max. 12 h 30 min) before the start of the first show jumping turn, respectively, 7 h 30 min (min 1 h 50 min, max. 13 h 0 min) before the second turn. Seven horses (35%) had access to hay waiting in the trailer between two turns. The statistical analysis revealed no significant influence of the concentrate feeding time point on lactate, triglyceride and insulin levels, but a significant influence on free fatty acids (FFA) and blood glucose concentrations. Roughage feeding of the show jumping horses 2-4 h prior to exercise revealed the most remarkable changes in blood parameters during the show jumping course. These results received under field conditions should be approved in future under standardized conditions.

Estimating energy losses with urine in the cat.

Urinary energy losses in cats have to be determined in energy balance trials as well as for the calculation of the metabolizable energy (ME) content of cat food. The aim of the present study was: first, to assess whether the energy content of cat urine quantified by bomb calorimetry differs from that quantified using GE (kJ) urine = 33 kJ × g C urine + 9 kJ × g N urine and investigate whether this difference could be attributed to influences of diets. Second, to assess whether the subtraction of 3.1 kJ/g of protein intake used for estimation of metabolizable energy content of cat foods is confirmed as usable. Data from 27 energy and protein balance trials from different studies with complete sampling of urine and faeces (29 cats in part A and 35 cats in part B) were used. Gross energy, carbon and nitrogen were determined in food, faeces and urine. Gross energy values in urine tended to be higher when determined with the formula of Hoffman and Klein compared to bomb calorimetry. The average relative difference of gross energy values between the methods was 18.8%. The mean energy loss in kJ/g of protein intake resulted in 3.7 kJ/g protein intake, which was not statistically significantly different (p = 0.12) from the tested value of 3.1 kJ/g of protein intake. In conclusion, the formula of Hoffman and Klein is not appropriate for the estimation of energy in cat urine. In balance studies, it is advisable to quantify the urinary energy content by bomb calorimetry. In the second part of the study, the protein correction factor to determine ME of 3.1 kJ/g protein intake for urinary energy losses of Kienzle et al. could be confirmed.

Overweight and impaired insulin sensitivity present in growing cats.

Obesity is a growing problem in pets as well as in humans. Overweight and obesity are linked to insulin sensitivity and subsequently in older cats, to an increased risk of developing diabetes mellitus. In the experimental cat population of the Institute of Animal Nutrition of the Vetsuisse Faculty, University of Zurich, an overweight phenotype in intact cats younger than 1 year became evident. The aims of the present study were to determine whether an association between insulin sensitivity and body condition score (BCS) or feline body mass index (FBMI) is already present during young adulthood in these cats and to test the hypothesis that the phenotype lean/overweight is significantly associated with monthly body weight during the growing period. Therefore, 41 kittens from the mentioned cat breeding colony were studied. They were weighed weekly and checked monthly (third to eighth month after birth) for BCS and FBMI. At the age of 8 months, they were classified into an overweight and lean phenotype based on BCS on a scale of 9 (median; maximum and minimum: overweight male (6.4; 6.8; 6.0); overweight female (6.1; 6.2; 6.0); lean male (5.4; 5.7; 5.0); lean female (5.2; 5.6; 5.0). A significant association between the phenotype and body weight was obvious during the growing period from the third to the 8 months (p = 0.0001). At month 8, body fat content was measured by dual energy X-ray absorptiometry and a glucose tolerance test to determine the insulin sensitivity index was performed. Insulin sensitivity was significantly associated with BCS (p = 0.0007) and body fat content (p < 0.0001) but not with sex (p = 0.61). Our data provide evidence that already in young intact cats; insulin insensitivity is significantly associated with BCS or a presumed phenotype lean/overweight.

A survey on the feeding of eventing horses during competition.

This study aims at the comparison of the actual feeding of horses with the recommendations from the literature, and it studies the effects of feeding and exercise on several blood metabolic parameters before and after exercise. Blood samples were collected from 25 horses during one-star eventing competitions and evaluated for blood glucose, insulin, lactate, free fatty acids and triglyceride levels. Questionnaires on the feeding practices of the horses were evaluated. The questionnaires revealed that during training, and on tournament days, horses received on average 4.3 kg of concentrate per day (min. 1.54 kg, max. 8 kg). The statistical analysis showed no significant effect of the amount of concentrate fed before exercise on the measured blood values. Oil was supplied as a supplementary energy source to 30% of the horses, but most of them only received very small quantities (0.02-0.4 l/day). Five horses (20%) had no access to salt supplements at all, and eleven horses (45%) had no access to salt on tournament days. Fifteen horses (60%) were supplied with mineral feed. Twenty-one horses (84%) had daily access to pasture during the training period. During competition, 55% of the horses received roughage ad libitum, compared with 37% during training. The majority of the horses received less roughage on days before the cross-country competition. It could not be ascertained whether feeding a large amounts of roughage had a beneficial effect on performance, because only a few horses in this study were fed with very restrictive roughage. Feeding of most of the horses was in agreement with the recommendations from the literature, except the need for sodium and chloride. The sodium and chloride need for sport horses may be overestimated in literature and needs to be re-evaluated.

Fermentation of six different forages in the semi-continuous fermentation technique Caesitec.

The aim of the present study was to compare carbohydrate degradation of forages which store carbohydrates either predominantly as fructan or starch, in horses' hindgut. The effects of an abrupt change from hay-based feeding to green fodder-based feeding on the caecal flora were tested with the in vitro hindgut simulation technique 'Caesitec'. Six trials with different forages (English ryegrass, tall fescue, grass mixture-horses, grass mixture-cows, lucerne, white clover) were conducted. During a 4-day stabilisation period, samples were taken once a day before loading the fermenters with hay. After diet-change to forage-based feeding, samples were taken four times a day. Ammonia and pH-value were measured before and 1, 2 and 6 h after loading the 'Caesitec'. Gas formation was measured daily. Bacterial numbers, lactate and short chain fatty acids were detected at four time-points of each trial. The grass mixtures contained the highest amounts of fructan. The pH-values were in the physiological range from pH 6 up to 7 (6.58-6.83) by feeding all forages. Gas formation, anaerobic and aerobic bacterial numbers increased after diet change from hay to any forage. The maximum amount of fructan (3.75 g/kg) in swiss pasture did not cause a permanent pathological change in the hindgut-flora.

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.

Activities of leukocyte enzymes in cats during feeding different diets.

The objective of the present study was to investigate the influence of diet composition on enzyme activities in feline leukocytes. In trial A, the enzyme activity was measured in six normal weight cats fed four cooked diets with varying protein, fat and carbohydrate contents and different protein sources. In trial B, six normal weight and six obese cats were fed on dry food of different protein and carbohydrate content. The enzyme activities did not show a distinct and consistent reaction to different diet compositions. Therefore, it is doubtful, if enzyme activities in feline leukocytes are an appropriate tool to examine the metabolic state or detect metabolic diseases in cats.

Toxocara eggs shed by dogs and cats and their molecular and morphometric species-specific identification: is the finding of T. cati eggs shed by dogs of epidemiological relevance?

Intestinal infections with Toxocara cati and Toxocara canis in their definitive host (felids and canids, respectively) are diagnosed by egg identification in faeces using coproscopical techniques. The Toxocara species is assumed to comply with the species from which the examined faeces were obtained, i.e. T. cati in cats and T. canis in dogs. We isolated and measured Toxocara eggs from faecal samples of 36 cats and 35 dogs from Switzerland and identified the Toxocara species by PCR. Amongst the isolates originating from dogs, 24 (68.5%) were determined as T. canis and 11 (31.5%) as T. cati. In all samples originating from cats, only T. cati was identified. Based on PCR identification, eggs of T. canis (n=241) and T. cati (n=442) were measured, revealing statistically significant different (p<0.001) mean sizes of 62.3 by 72.7 µm for T. cati and 74.8 by 86.0 µm for T. canis eggs. Considering that coprophagy is not unusual for dogs, a considerable percentage of Toxocara infections coproscopically diagnosed in dogs, as well as assumptions on anthelminthic resistance in regularly treated dogs, might in fact relate to intestinal passages of eggs following the uptake of other animals' faeces.

Evaluation of a novel real-time continuous glucose-monitoring system for use in cats.

BACKGROUND: The Guardian REAL-Time is a continuous glucose-monitoring system (CGMS) recently developed to provide instantaneous interstitial glucose concentrations; the system does not require a monitor being fixed to the animal. HYPOTHESIS: The CGMS provides accurate and reproducible real-time readings of glucose concentration in cats. ANIMALS: Thirty-two diabetic cats, 2 cats with suspected insulinoma, and 5 healthy cats. METHODS: Prospective, observational study. CGMS accuracy was compared with a reference glucose meter at normal, high, and low blood glucose concentrations using error grid analysis. Reading variability of 2 simultaneously used CGMS was determined in diabetic cats by calculating correlation and percentage of concordance of paired data at different glycemic ranges. The time interval between increasing glycemia and a rise in interstitial fluid glucose measured by the CGMS was assessed in healthy cats receiving glucose IV; the time point of maximal increase in interstitial glucose concentrations was calculated. RESULTS: The CGMS was 100, 96.1, and 91.0% accurate at normal, high, and low blood glucose concentrations. Measurements deviated from reference by -12.7 +/- 70.5 mg/dL at normal, -12.1 +/- 141.5 mg/dL at high, and -1.9 +/- 40.9 mg/dL at low glucose concentrations. Overall, paired CGMS readings correlated significantly (r = 0.95, P < .0001) and concordance was 95.7%. The median delay after IV administration of glucose to an increase in interstitial glucose was 11.4 minutes (range: 8.8-19.7 minutes). CONCLUSIONS AND CLINICAL IMPORTANCE: Although some readings substantially deviated from reference values, the CGMS yields reproducible results, is clinically accurate in cats with hyperglycemia and euglycemia, and is slightly less accurate if blood glucose concentrations are low. Rapidly increasing interstitial glucose after a glycemic rise suggests that the CGMS is suitable for real-time measurement under clinical conditions.

[Puppy feeding in Switzerland]. / Welpenfütterung in der Schweiz.

In this study breeders and owners of 8 different dog breeds (Beagle, Bernese Mountain Dog, Cavalier King Charles Spaniel, Great Dane, German Shepherd (GS), Labrador, Papillon, Sheltie) were interviewed to obtain information on puppy feeding in Switzerland. Besides answering a questionnaire (husbandry and feeding of the puppies), the participation in this study included weekly weighing of the animals as well as exact documentation of the amount fed to the animals. Totally 67 dog breeders and 131 new owners of puppies participated. The weight development of the puppies was mostly parallel to the growth curve in the GS, Labradors and Shelties. There were some substantial differences to the ideal growth curve within the other breeds. The daily mean energy requirement was estimated too high, when including the growth curves. 80 - 90 % of the recommendations would be sufficient for most animals. The calcium supply was in the range of tolerance in all breeds. Nearly all breeders used commercially available complete food while raising the puppies. No breed-specific differences could be shown.

Energy and protein needs of cats for maintenance, gestation and lactation.

In the present investigation, data on the energy intakes and energy needs, as well as protein and fat accretion, of queens during pregnancy, during lactation and after lactation are given. Eleven adult cats were used as experimental animals. Data were collected during the fourth and seventh week of pregnancy, the second and sixth week of lactation and the second and sixth week after lactation. The cats were fed dry kitten food. During gestation and after lactation, all measurements were performed with respiration chambers. During lactation, balance trials without respiration chambers were performed. Body weight was measured and nitrogen, carbon and energy balances were calculated. From these, protein and fat accretion, as well as the metabolisable energy intake, was calculated. The weight gain during gestation was linearly independent of the number of kittens. During lactation, all cats lost weight; nevertheless, all cats except one were heavier 2 weeks after lactation than at mating. The energy intake of the cats during gestation was 1.8 times the maintenance requirement in the fourth week and two times maintenance requirement in the seventh week, and these energy intakes differed greatly among individuals. The energy intake of the cats during lactation was clearly higher than that recommended by National Research Council (NRC)(1), whereas the recommended protein intake in the second week of lactation was met. As the calculated protein balance was negative, the NRC recommendation for protein intake seems to be too low. In comparison to previous data, the cats showed a higher energy intake during lactation (median 502kJ/kgBW/d, second week lactation), and the weight loss was much lower. Further investigations on pregnant and lactating cats are necessary to complete the database.

Mineral content of hay harvested in Bavarian and Swiss horse farms. Predictive value of cutting time, number of cut, botanical composition, origin and fertilization.

Hay samples from 29 horse farms in Southern Upper Bavaria and 31 horse farms in Switzerland were taken and analysed for minerals such as calcium, magnesium, phosphorus and potassium. The content of herbs and legumes in the hay was determined and the Weende analysis was performed. Comparison between Bavaria and Switzerland resulted in comparable calcium contents of approximately 4 g Ca/kg dry matter (DM) in grass hay. Hay with more than 10% of herbs and legumes found only in Switzerland showed higher Ca contents of >7 g Ca/kg DM in hay. The mineral contents of phosphorus and potassium were lower in hay from Switzerland (P: 1.8 +/- 0.7 g/kg DM, K: 15.4 +/- 5.1 g/kg DM) than from Bavaria (P: 3.8 +/- 0.6 g/kg DM, K: 20.0 +/- 6.0 DM), whereas the magnesium content of the hay showed no difference between the regions ( approximately 1.5 g Mg/kg DM). Very late first grass hay cuts showed low magnesium and calcium (<4 g/kg DM) contents. Further minerals in the hay for horses differed from those in dairy cattle in the same region. Fertilization showed only small effects on the mineral contents of the hay produced for horses. Therefore, in calculation of ration and production of supplements for horses, these differences should be taken into account.

Judgement of hygienic quality of roughage in horse stables in Switzerland.

In 46 horse farms all over Switzerland, the hygienic quality of the roughage (including silages) was investigated. Therefore, a macroscopic examination as well as the microbial counts (bacteria, yeasts and moulds) was carried out. Further, the contents of lipopolysaccharides (LPS) and the contamination with deoxynivalenol (DON) were determined. In all roughages, the dry matter (DM) content was measured and in silages additionally the pH was measured. Predominantly, the straw showed a lower hygienic quality than hay and silages. The LPS contents were significantly higher in straw than those in hay samples. The macroscopic examination and the microbial counts showed a tendency in the same direction. Eight straw samples and one hay sample with DON contamination were found. The silages showed a DM content with a median of 65.8%. The pH of those silages was between 4.3 and 5.9. Despite the high values of DM and pH, the silages showed mainly a high hygienic quality. In total, the hygienic standard of straw was worse than that of hay. This should be taken into account also in straw used as bedding as much as the hygienic quality of hay.

Effect of vitamin E supplementation of sheep and goats fed diets supplemented with polyunsaturated fatty acids and low in Se.

Vitamin E (VitE) and selenium (Se) are an essential part of the antioxidative functions of metabolism. There are situations of low supply of both micronutrients. As VitE is involved in ruminal biohydrogenation of polyunsaturated fatty acids (PUFA) and their protection against oxidation in metabolism, diets supplemented with PUFA may challenge VitE to an extent making recommended supplies insufficient. Twelve goats and sheep each were fed a diet supplemented with PUFA and characterised by low Se and limited VitE contents during the last 2 months of gestation and the first 2 months of lactation. The basal diet consisted of hay and concentrate. Six goats and sheep received extra VitE, while the control groups received no extra VitE. Blood and milk samples were taken. In addition, liver, heart muscle and spleen samples were obtained from the offspring after slaughtering at an age of 8 weeks. No significant changes were observed in serum Se and VitE. A significant increase in serum VitE concentrations between 2 and 4 weeks postpartum (pp) was evident in the supplemented kids. In 4, 6 and 8 weeks pp, the serum concentrations of VitE in the supplemented kids were significantly higher compared to the unsupplemented group. In the kids, VitE was higher in liver of the supplemented groups. There were no significant differences in response to extra VitE between sheep and goat. The kids responded to serum VitE different from that of lambs, as a significant difference was observed between supplemented and unsupplemented animals in the goat kids, but not the lambs. In conclusion, goats and sheep have to be viewed differently and may not be considered alike relating to VitE/Se metabolism and requirements, especially in young animals.

Additional data on energy requirements of young adult cats measured by indirect calorimetry.

Measurements of nitrogen, carbon and energy balances were done on eight female adult cats aged approximately 1.5 years with the help of respiration chambers. The cats were fed with a marked dry food for kittens (Biomill kitten). Faeces and urine samples were collected with the help of a special cat toilet that allows the single collection of both materials. The calculated energy requirement of these eight female cats was 239.6 kJ ME/kg BW/day (R(2) = 0.98). Including the data, on 12 young male adult cats, of Läuger, (2001), an energy requirement of 238 kJ ME/kg BW/day (R(2) = 0.95) could be calculated. From these data, it can be followed that the energy requirements of young, active cats are higher than that of the older cats. The method of indirect calorimetry does not necessarily lead to lower energy requirements.

[Nutrient content and adequacy of roughage for horse nutrition: an investigation under Swiss conditions]. / Nährstoffgehalte und Eignung des Grundfutters zur Pferdefütterung: Eine Erhebung unter schweizerischen Bedingungen.

For horses no special tables related to nutrients for Swiss roughage exist. For this reason samples of hay, straw, silage/haylage and green forage were taken from 46 horse keeping farms in 22 cantons. The samples were judged by sense and following the nutrient--and macromineral--content as well as the content of fructans were analysed. Regarding its quality no sample was totally inadequate for horses. The mean contents of crude protein in Swiss hay for horses were clearly lower than in hay for ruminants and in German hay for horses. The mineral contents (calcium, magnesium, phosphorus) showed a wide range. On average they were also lower than the values provided in tables for ruminants. Except for one sample the silages/haylages showed a dry matter content of more than 40 %. No nutrient tables for silage or haylage, which are such high in dry matter contents, were found in the literature. The contents offructans in silage/haylage and green forage also showed a wide range with a maximum of 94 g/kg DM fructan.

Mutations within the FGF5 gene are associated with hair length in cats.

Hereditary hair length variability in mice and dogs is caused by mutations within the fibroblast growth factor 5 (FGF5) gene. The aim of this study was to evaluate the feline FGF5 orthologue as a functional candidate gene for the long hair phenotype in cats, which is recessive to short hair. We amplified the feline FGF5 cDNA and characterised two alternatively spliced transcripts by RT-PCR. Comparative cDNA and genomic DNA sequencing of long- and short-haired cats revealed four non-synonymous polymorphisms in the FGF5 coding sequence. A missense mutation (AM412646:c.194C>A) was found in the homozygous state in 25 long-haired Somali, Persian, Maine Coon, Ragdoll and crossbred cats. Fifty-five short-haired cats had zero or one copy of this allele. Additionally, we found perfect co-segregation of the c.194C>A mutation within two independent pedigrees segregating for hair length. A second FGF5 exon 1 missense mutation (AM412646:c.182T>A) was found exclusively in long-haired Norwegian Forest cats. The c.182T>A mutation probably represents a second FGF5 mutation responsible for long hair in cats. In addition to the c.194C>A mutation, a frameshift mutation (AM412646:c.474delT) was found with a high frequency in the long-haired Maine Coon breed. Finally, a missense mutation (AM412646:c.475A>C) was also associated with the long-haired phenotype in some breeds. However, as one short-haired cat was homozygous for this polymorphism, it is unlikely that it has a functional role in the determination of hair length.