Genetic variability among and within domestic Old and New World camels at the α-lactalbumin gene (LALBA) reveals new alleles and polymorphisms responsible for differential expression.

α-Lactalbumin (α-LA), which is encoded by the LALBA gene, is a major whey protein that binds to Ca2+ and facilitates lactose synthesis as a regulatory subunit of the synthase enzyme complex. In addition, it has been shown to play central roles in immune modulation, cell-growth regulation, and antimicrobial activity. In this study, a multitechnical approach was used to fully characterize the LALBA gene and its variants in both coding and regulatory regions for domestic camelids (dromedary, Bactrian camel, alpaca, and llama). The gene analysis revealed a conserved structure among the camelids, but a slight difference in size (2,012 bp on average) due to intronic variations. Promoters were characterized for the transcription factor binding sites (11 found in total). Intraspecies sequence comparison showed 36 SNPs in total (2 in the dromedary, none in the Bactrian camel, 22 in the alpaca, and 12 in the llama), whereas interspecies comparison showed 86 additional polymorphic sites. Eight SNPs were identified as trans-specific polymorphisms, and 2 of them (g.112A>G and g.1229A>G) were particularly interesting in the New World camels. The first creates a new binding site for transcription factor SP1. An enhancing effect of the g.112G variant on the expression was demonstrated by 3 independent pGL3 gene reporter assays. The latter is responsible for the p.78Ile>Val AA replacement and represents novel allelic variants (named LALBA A and B). A link to protein variants has been established by isoelectric focusing (IEF), and bioinformatics analysis revealed that carriers of valine (g.1229G) have a higher glycosylation rate. Genotyping methods based on restriction fragment length polymorphism (PCR-RFLP) were set up for both SNPs. Overall, adenine was more frequent (0.54 and 0.76) at both loci. Four haplotypes were found, and the AA and GA were the most common with a frequency of 0.403 and 0.365, respectively. Conversely, a putative biological gain characterizes the haplotype GG. Therefore, opportunities for rapid directional selection can be realized if this haplotype is associated with favorable milk protein properties. This study adds knowledge at the gene and protein level for α-LA (LALBA) in camelids and importantly contributes to a relatively unexplored research area in these species.

A novel allelic donkey ß-lactoglobulin I protein isoform generated by a non-AUG translation initiation codon is associated with a nonsynonymous SNP.

ß-lactoglobulin I (ß-LG I) is one of the most important whey proteins in donkey milk. However, to our knowledge, there has been no study focusing on the full nucleotide sequences of this gene (BLG I). Current investigation of donkey BLG I gene is very limited with only 2 variants (A and B) characterized so far at the protein level. Recently, a new ß-LG I variant, with a significantly higher mass (+1,915 Da) than known variants has been detected. In this study, we report the whole nucleotide sequence of the BLG I gene from 2 donkeys, whose milk samples are characterized by the ß-LG I SDS-PAGE band with a normal electrophoretic mobility (18,514.25 Da, ß-LG I B1 form) the first, and by the presence of a unique ß-LG I band with a higher electrophoretic mobility (20,428.5 Da, ß-LG I D form) the latter. A high genetic variability was found all over the 2 sequenced BLG I alleles. In particular, 16 polymorphic sites were found in introns, one in the 5' flanking region, 3 SNPs in the 5' untranslated region and one SNP in the coding region (g.1871G > A) located at the 40th nucleotide of exon 2 and responsible for the AA substitutions p.Asp28 > Asn in the mature protein. Two SNPs (g.920-922CAC > TGT and g.1871G/A) were genotyped in 93 donkeys of 2 Italian breeds (60 Ragusana and 33 Amiatina, respectively) and the overall frequencies of g.920-922CAC and g.1871A were 0.3065 and 0.043, respectively. Only the rare allele g.1871A was observed to be associated with the slower migrating ß-LG I. Considering this genetic diversity and those found in the database, it was possible to deduce at least 5 different alleles (BLG I A, B, B1, C, D) responsible for 4 potential ß-LG I translations. Among these alleles, B1 and D are those characterized in the present research, with the D allele of real novel identification. Haplotype data analysis suggests an evolutionary pathway of donkey BLG I gene and a possible phylogenetic map is proposed. Analyses of mRNA secondary structure showed relevant changes in the structures, as consequence of the g.1871G > A polymorphism, that might be responsible for the recognition of an alternative initiation site providing an additional signal peptide. The extension of 19 AA sequence to the mature protein, corresponding to the canonical signal peptide with an additional alanine residue, is sufficient to provide the observed molecular weight of the slower migrating ß-LG I encoded by the BLG I D allele.

Sequencing of lipoprotein lipase gene in the Mediterranean river buffalo identified novel variants affecting gene expression.

Lipoprotein lipase (LPL) is a key enzyme for lipid metabolism, playing a fundamental role in the composition of fat in adipose tissue and milk. The LPL gene has been seldom investigated in dairy ruminants and barely studied in river buffalo (Bubalus bubalis). The aim of this work was to explore the genetic diversity of LPL and its promoter and to identify functional mutations, using a combined approach based on sequencing, dual-color electrophoretic mobility shift assay, and quantitative PCR. Thirteen consensus sequences for transcription factors were found in the promoter. Eleven SNP were detected, and the attention was focused on the SNP with potential functional effects: g.-446A>G, because the presence of G created a consensus motif for the transcription factor Sp1, and g.107A>G, which was the only exonic SNP. We developed PCR-RFLP methods for genotyping the 2 SNP and calculated the allele frequencies. A strong linkage disequilibrium (D' = 1; r2 = 0.903) was found between the 2 SNP. The dual-color electrophoretic mobility shift assay demonstrated that only genotype g.-446GG allowed the binding of the Sp1 transcription factor, resulting in overexpression of the gene (~2.5 fold), as confirmed by the quantitative PCR results. Haploinsufficiency is proposed as a regulation mechanism. This study adds further knowledge on the structure of the LPL gene and its expression in river buffalo, with potential effects on milk qualitative and quantitative production.

The single nucleotide polymorphism g.133A>C in the stearoyl CoA desaturase gene (SCD) promoter affects gene expression and quali-quantitative properties of river buffalo milk.

The stearoyl-CoA desaturase (SCD) gene has been investigated in depth in ruminants because of its effect on milk fat composition. In river buffalo, the single nucleotide polymorphism (SNP) g.133A>C in the gene promoter has been associated with milk quality and yield. However, the biological reason for such effects remains unexplored. In this study, we combined mRNA profile analysis, an electromobility shift assay, and quantitative PCR to elucidate the role of this SNP on gene transcription and its effects on milk fat traits. A preliminary genotyping of g.133A>C was carried out on a group of 303 river buffaloes to choose individuals for the downstream applications. Analysis of allele frequencies showed an increase in the minor allele C (0.25) compared with previous findings (0.16). Six animals (2 for each genotype) were chosen for cloning and 216 positive cDNA recombinant clones for SCD (72 per genotype) were analyzed by PCR. All clones showed the same length on agarose gel; therefore, random clones were chosen for sequencing. No qualitative differences were found and all gene transcripts assembled correctly. An electrophoretic mobility shift assay was performed to evaluate the binding of the transcription factor Sp1 to DNA sequences including g.133A>C. Genotype CC showed a higher binding (mean ± standard error of the mean) than genotype AA in 2 different conditions [Enzo buffer (EB), Enzo Life Science Inc., Farmingdale, NY: 201.77 ± 4.06 vs. 141.65 ± 3.77 band intensity values and Poletto buffer (PB): 95.90 ± 1.15 vs. 67.30 ± 2.14 band intensity values]. The subsequent quantitative PCR confirmed the upregulation of the CC genotype compared with the AA and AC genotypes. The association study with milk fat traits revealed a favorable effect of allele C. The heterozygous genotype had the highest values for monounsaturated fatty acids, oleic acid (C18:1 cis-9), polyunsaturated fatty acids, and odd- and branched-chain fatty acids, and the lowest values for saturated fatty acids and atherogenic and thrombogenic indices; the heterozygous genotype differed significantly from the AA genotype. The AC genotype has previously been associated with higher milk yield. Therefore, the g.133A>C SNP is a marker with dual effects and is an interesting candidate for assisted selection programs in river buffalo. These data clarified the biological role of the SNP g.133A>C in the SCD promoter and how it affects gene function, providing important knowledge on the genetic background of lipid metabolism, including the future possibility of selecting alleles with quantitatively or qualitatively favorable effects.

Remarkable genetic diversity detected at river buffalo prolactin receptor (PRLR) gene and association studies with milk fatty acid composition.

Prolactin is an anterior pituitary peptide hormone involved in many different endocrine activities and is essential for reproductive performance. This action is mediated by its receptor, the prolactin receptor, encoded by the PRLR gene. In this study, we sequenced and characterized the Mediterranean river buffalo PRLR gene (from exon 3 to 10), and we found remarkable genetic diversity. In particular, we found 24 intronic polymorphisms and 13 exonic SNPs, seven of which were non-synonymous. Furthermore, the polymorphisms identified in the 3'-UTR were investigated to establish their possible influence on microRNA binding sites. Considering all the amino acid changes and the observed allelic combinations, it is possible to deduce at least six different translations of the buffalo prolactin receptor and, consequently, the presence at the PRLR gene of at least six alleles. Furthermore, we identified a deletion of a CACTACC heptamer between nucleotides 1102 and 1103 of exon 10 (3'-UTR), and we developed an allele-specific PCR to identify the carriers of this genetic marker. Finally, the SNP g.11188A>G, detected in exon 10 and responsible for the amino acid replacement p.His328Arg, was genotyped in 308 Italian Mediterranean river buffaloes, and an association study with milk fat traits was carried out. The statistical analysis showed a tendency that approached significance for the AA genotype with higher contents of odd branched-chain fatty acids. Thus, our results suggest that the PRLR gene is a good candidate for gene association studies with qualitative traits related to buffalo milk production.

Lama glama αS1-casein: Identification of new polymorphisms in the CSN1S1 gene.

South American camelids have been poorly genetically investigated and little information is available in llamas (Lama glama) regarding the diversity of the caseins at the protein and gene level. Exon skipping and duplication events previously reported in the αS1-casein gene (CSN1S1) led us to investigate the genetic variability at this locus. Seventy-two positive clones for the αS1-casein transcripts were analyzed and randomly sequenced. The comparative analysis of the sequences revealed 2 transitions, c.366A>G and c.690T>C, at the 10th nucleotide of exon 12 and 94 bp of exon 19, respectively. These SNP are responsible for 2 amino acid changes, Ile→Val in position 86 and Tyr→His in position 194 of the mature protein. Both polymorphisms clarify the genetic events behind the protein variants A and B. This result was confirmed by isoelectric focusing analysis of llama milk samples. Quick methods based on PCR-RFLP and allele-specific PCR were set up for allelic discrimination in a population of 128 animals. Based on genotyping results, 4 haplotypes were observed and the estimated frequencies indicated B as the most common haplotype (0.629) in the investigated population. These data add knowledge to the genetic variability of a species little investigated, and open opportunity for new investigation in the field of milk protein for South American camelids, including the possibility, in the future, to select alleles with favorable characteristics.

Transcript analysis at DGAT1 reveals different mRNA profiles in river buffaloes with extreme phenotypes for milk fat.

Buffalo DGAT1 (diacylglycerol O-acyltransferase 1) was mainly investigated for the characterization of the gene itself and for the identification of the K232A polymorphism, similar to what has been accomplished in cattle, although no information has been reported so far at the mRNA level. The importance of DGAT1 for lipid metabolism led us to investigate the transcript profiles of lactating buffaloes characterized as high (9.13 ± 0.23) and low (7.94 ± 0.29) for milk fat percentage, and to explore the genetic diversity at the RNA and DNA level. A total of 336 positive clones for the DGAT1 cDNA were analyzed by PCR and chosen for sequencing according to the differences in length. The clone assembling revealed a very complex mRNA pattern with a total of 21 transcripts differently represented in the 2 groups of animals. Apart from the correct transcript (17 exons long), the skipping of exon 12 is the most significant in terms of distribution of clones with 11.6% difference between the 2 groups, whereas a totally different mRNA profile was found in approximately 12% of clones. The sequencing of genomic DNA allowed the identification of 10 polymorphic sites at the intron level, which clarify, at least partially, the genetic events behind the production of complex mRNA. Genetic diversity was found also at the exon level. The single nucleotide polymorphism c.1053C>T represents the first example of polymorphism in a coding region for the DGAT1 in the Italian Mediterranean breed. To establish whether this polymorphism is present in other buffalo breeds, a quick method based on PCR-RFLP was set up for allelic discrimination in the Italian Mediterranean and the Romanian Murrah (200 animals in total). The alleles were equally represented in the overall population, whereas the analysis of the 2 breeds showed different frequencies, likely indicating diverse genetic structure of the 2 breeds. The T allele might be considered as the ancestral condition of the DGAT1 gene, being present in the great part of the sequenced species. These data add knowledge at the transcript and genetic levels for the buffalo DGAT1 and open the opportunity for further investigation of other genes involved in milk fat metabolism for the river buffalo, including the future possibility of selecting alleles with quantitative or qualitative favorable effects (or both).

The SNP g.1311T>C associated with the absence of ß-casein in goat milk influences CSN2 promoter activity.

Quantitative individual differences in the amount of ß-casein in goat milk are determined by at least nine alleles. In particular, two alleles (CSN2(0) and CSN2(01) ) are associated with an undetectable amount of this protein in milk. The CSN2(01) allele is characterized by a single nucleotide substitution at position 373 of the seventh exon (AJ011018:g.8915C>T), responsible for the formation of a premature stop codon at the 182 position. Herein, we report the contribution of the SNP g.1311T>C, which demonstrates a linkage with the SNP AJ011018:g.8915C>T, to the promoter transcriptional activity. Particularly, we indicate that the nucleotide C at position 1311 negatively affects the promoter activity of the CSN2 gene.

Short communication: Translational efficiency of casein transcripts in Mediterranean river buffalo.

Buffalo milk is characterized by the presence of all 4 casein fractions (α(S1), ß, α(S2), and κ) encoded by the 4 tightly linked autosomal genes (CSN1S1, CSN2, CSN1S2, and CSN3, respectively). In the present paper, we report for the first time a quantitative characterization of buffalo casein transcripts and show that the 4 genes are not transcribed and translated with the same efficiency. In particular, the analysis of individual milk samples obtained from 9 Mediterranean river buffaloes showed that the most abundant casein fractions were ß (53.45%) and α(S1) (20.61%), followed by α(S2) and κ, at 14.28 and 11.66%, respectively. Quantification of the corresponding mRNA showed that the percentage of transcripts of the 4 caseins was 16.48, 23.18, 55.87, and 4.47% for α(S1), ß, α(S2), and κ, respectively. Translation efficiency was 0.25 for CSN1S2, 1.31 for CSN1S1, 2.39 for CSN2, and 2.69 for the CSN3 transcripts, respectively. A comparison of nucleotide sequences with the Kozak consensus sequence was also carried out to investigate if the mRNA sequences might be responsible for the observed differences.

Peritoneal washing cytology in gastric cancer. How, when and who will get a benefit? A review.

The most frequent cause of treatment failure following surgery for gastric cancer is peritoneal metastasis. The ability to predict the likelihood of peritoneal recurrence should improve the therapeutic approach to gastric cancer. Cytological analysis of peritoneal washings is thought to be useful for direct detection of free cancer cells in the peritoneal cavity. Intraperitoneal free cancer cells (IFCC) isolated during peritoneal washing in patients with gastric cancer, have been demonstrated to be significantly and independently related to the prognosis, influencing both early recurrence and poor survival, so that since 1998 the Japanese Classification of Gastric Carcinoma (JCGC) recommend peritoneal wash cytology (PWC) for the local staging. In Western countries PWC is not uniform practice, because of several controversies regarding the low sensitivity rate of conventional cytology, the correct application of molecular diagnosis (immunostaining and RT_PCR) and the exact role of PWC in the clinical practice. The authors examine the current apply of peritoneal washing in gastric cancer, emphasizing the clinical implication of peritoneal cytology by analyzing the different modality and techniques to perform it (conventional cytology, immunocytochemistry, RT-PCR), when to achieve it during the diagnostic or clinical work-up (at the staging or during the surgical treatment), and who will get a benefit (all patients or selected patients).

Sequence analysis and genetic variability of stearoyl CoA desaturase (SCD) gene in the Italian Mediterranean river buffalo.

Stearoyl-CoA desaturase (SCD) plays a key metabolic role by changing the saturated FA content of ruminant milk and meat. In this study we characterized for the first time the stearoyl-CoA desaturase (SCD) gene in river buffalo (Bubalus bubalis) and investigated its genetic variability. On a total of 78 buffaloes, 15 SNPs were detected and 6 of them were preliminarily genotyped. In particular, the g.133A>C SNP was found to create a new consensus site for the SP1 binding site, thus generating a new tandem repeat in the promoter region. A preliminary association study with the milk fatty acid content showed that the C allele significantly affects the total desaturation index (P<0.01). Linkage disequilibrium analysis allowed identification of 7 haplotypes and 4 tag SNPs. Such polymorphisms could represent useful genetic markers for association studies with fatty acid composition, but further studies are needed to evaluate their potential use to improve the nutritional quality of the dairy products.

X-Y sperm aneuploidy in 2 cattle (Bos taurus) breeds as determined by dual color fluorescent in situ hybridization (FISH).

The present study was undertaken to investigate aneuploidy rates in the sperm populations of 2 cattle (Bos taurus) breeds by using dual color fluorescent in situ hybridization (FISH) with Xcen and Y chromosome-specific painting probes, obtained by chromosome microdissection and DOP-PCR. Frozen semen from 10 Italian Friesian and 10 Italian Brown testing bulls was used for the investigation. For each bull, more than 5,000 sperm were analyzed, for a total of 52,586 and 51,342 sperm cells for the 2 breeds, respectively. The present study revealed - in both breeds - a preponderance of the Y-bearing sperm compared to the X-bearing sperm. Within each breed, a statistically significant variation in the various classes of aneuploidy (XX, YY and XY) was found: differences were found in the Friesian breed among the 3 diploidy classes, and in the Brown breed, among the 3 disomy classes (p < 0.05) as well as among the 3 diploidy classes (p < 0.01). However, the 2 breeds did not differ significantly in the overall mean rates of X-Y aneuploidy (disomy + diploidy) which amounts to 0.162% in the Italian Friesian and 0.142% in the Italian Brown. When meiosis I (MI) and II (MII) errors were compared, statistically significant differences (p < 0.01) were found in the disomy classes and in both breeds, whereas the differences between diploidy classes were not significant. Compared to humans, a lower level of aneuploidy has been found in the domestic species analyzed so far. The present study contributes to the establishment of a baseline level of aneuploidy in the sperm populations of 2 cattle breeds which could be used for monitoring future trends of reproductive health, especially in relation to environmental changes and mutagens.

Chromosomal expression and localization of aphidicolin-induced fragile sites in the standard karyotype of river buffalo (Bubalus bubalis).

The present study reports on the chromosomal expression and localization of aphidicolin-induced fragile sites in the standard karyotype of river buffalo (Bubalus bubalis, 2n = 50) with the aim of establishing a 'fragile site map' of the species. Totally, 400 aphidicolin-induced breakages were analyzed from eight young and clinically healthy animals, four males and four females; these breakages were localized in 106 RBG-negative chromosome bands or at the band-interband regions. The number of breakages per chromosome did not vary statistically 'among' the animals investigated but the differences among individual chromosomes were highly significant thus indicating that the chromosomal distribution of the breakages is not random and appears only partially related to chromosome length. Fragile sites were statistically determined as those chromosomal bands showing three or more breakages. In the river buffalo karyotype, 51 fragile sites were detected and localized on the standardized ideogram of the species. The most fragile bands were as follows: 9q213 with 24 breakages out of 400; 19q21 with 16, 17q21 and inacXq24 with 15, 15q23 with 13 and 13q23 with 12 breaks, respectively. Previous gene mapping analysis in this species has revealed that the closest loci to these fragile sites contain genes such as RASA1 and CAST (9q214), NPR3 and C9 (19q19), PLP and BTK (Xq24-q25), OarCP09 (15q24), and EDNRB (13q22) whose mutations are responsible for severe phenotypic malformations and immunodeficiency in humans as well as in mice and meat quality in pigs. Further cytogenetic and molecular studies are needed to fully exploit the biological significance of the fragile sites in karyotype evolution of domestic animals and their relationships with productive and reproductive efficiency of livestock.

[Teaching in the operative room: the benefit of daysurgery on surgical trainees]. / Attività didattica in sala operatoria: utilità della day-surgery nell'iter formativo dello specializzando in chirurgia.

AIM: Aim of the study was to evaluate the operative time and the incidence of post-operative complications in a group of patients undergoing Lichtenstein inguinal hernia repair performed either by surgical residents or senior surgeons in a day-surgery setting. PATIENTS AND METHODS: The study population consisted of 198 patients: group I (n=102), in which the operator was a senior surgeon, group II (n=96), in which the operator was a resident supervised by a senior surgeon. We recorded the duration of the operation and the complications following the procedure, and statistically compared them between group I and II. RESULTS: Our analysis showed that there was a statistically significant difference between the two groups only for the mean operative time, being shorter in group I (62 vs 82 min, p>0.05), while no significant difference was found for the incidence of complications. CONCLUSION: In conclusion, the day-surgery setting allows a high quality training of young surgeons, based on performing minor surgical procedures such has inguinal hernia repair. This training allows a step by step supervised learning process that does not jeopardize the efficacy of the treatment as well as the patient safety. The major cost due to the increase in operative time should be considered as an investment in young surgeons education.

An SNP in the goat CSN2 promoter region is associated with the absence of beta-casein in milk.

So far, at least eight alleles in the goat CSN2 locus have been associated with the level of beta-casein expression in milk. Alleles CSN2(A), CSN2(A1), CSN2(B), CSN2(C), CSN2(D) and CSN2(E) have been associated with normal content (allele effects of about 5 g of beta-casein per litre), whereas the CSN2(0) and CSN2(01) alleles have been associated with non-detectable levels of beta-casein. Most of these alleles have been characterized genetically. Herein, we report the identification of a previously unreported SNP in the goat CSN2 promoter region (AJ011018:g.1311T>C), which is associated with the absence of beta-casein in the milk. Furthermore, we developed a PCR-based method that allows detection of this mutation.

Cytogenetic characterization of alpaca (Lama pacos, fam. Camelidae) prometaphase chromosomes.

The present study provides specific cytogenetic information on prometaphase chromosomes of the alpaca (Lama pacos, fam. Camelidae, 2n = 74) that forms a basis for future work on karyotype standardization and gene mapping of the species, as well as for comparative studies and future genetic improvement programs within the family Camelidae. Based on the centromeric index (CI) measurements, alpaca chromosomes have been classified into four groups: group A, subtelocentrics, from pair 1 to 10; group B, telocentrics, from pair 11 to 20; group C, submetacentrics, from pair 21 to 29; group D, metacentrics, from pair 30 to 36 plus sex chromosomes. For each chromosome pair, the following data are provided: relative chromosome length, centromeric index, conventional Giemsa staining, sequential QFQ/C-banding, GTG- and RBG-banding patterns with corresponding ideograms, RBA-banding and sequential RBA/silver staining for NOR localization. The overall number of RBG-bands revealed was 391. Nucleolus organizer-bearing chromosomes were identified as pairs 6, 28, 31, 32, 33 and 34. Comparative ZOO-FISH analysis with camel (Camelus dromedarius) X and Y painting probes was also carried out to validate X-Y chromosome identification of alpaca and to confirm close homologies between the sex chromosomes of these two species.

Tibial plateau levelling osteotomy in 69 small breed dogs using conically coupled 1.9/2.5 mm locking plates. A clinical and radiographic retrospective assessment.

OBJECTIVE: To report clinical experiences with the tibial plateau levelling osteotomy (TPLO) procedure in small breed dogs with cranial cruciate ligament (CCL) disease using specific, conically coupled, 1.9/2.5 mm locking plates and evaluating short-term complications and outcome. METHODS: Medical records of small breed dogs (<15 kg) that underwent TPLO using 1.9/2.5 mm locking plates were reviewed retrospectively. The preoperative, postoperative and six to eight weeks postoperative tibial plateau angle (TPA) measurements were determined from the radiographic images. Lameness evaluation was assessed subjectively preoperatively and six to eight weeks postoperatively. RESULTS: Sixty-nine small breed dogs (n = 79 stifles) were included in the study. Mean (± SD) preoperative TPA was 29.0 ± 3.4°, postoperative TPA was 5.8 ± 2.5°, and six to eight weeks postoperative TPA was 7.3 ± 4.1°. Sixteen complications occurred in 12 out of 79 TPLO procedures: three were intra-operative (intra-articular screw placement) and 13 were postoperative complications, of which nine were identified as minor complications not requiring surgical reintervention, and four as major complications requiring additional surgical intervention, including tibial tuberosity fracture (n = 1), osteomyelitis (n = 1), screw failure (n = 1), and plate breakage (n = 1). Lameness scores by clinical assessment reduced from a median value of 3/4 preoperatively to 1/4 at six to eight weeks postoperatively. CLINICAL SIGNIFICANCE: 1.9/2.5 mm locking plates appear to be a valid choice of implant for the stabilization of unilateral TPLO in small breed dogs.

Development and evaluation of a fermented cabrito snack stick product.

In order for the meat goat producer to survive, new avenues for marketing goats must be created. Currently, the live animal is sold directly to consumers, or to brokers who in turn sell the animal directly to consumers or retail stores that cater to various ethnic groups. The production of value-added products with appeal to North American consumers, as well as current ethnic consumers, should result in increased profitability of the meat goat. The objectives of this study were to develop a value added product, cabrito snack sticks, using goat meat as the sole meat ingredient; evaluate soy protein concentrate (SPC) at various levels in an effort to reduce the cost of the product; determine consumer acceptability of the product; and conduct a cost analysis to determine the approximate market price for the product. Three fermented cabrito snack stick products were manufactured containing either 0, 1.75 or 3.50% SPC and stored at 2±1°C until evaluated. The snack sticks were evaluated for sensory characteristics, proximate analysis, pH, water activity and smokehouse yields. Trained panelists detected no significant flavor differences (P>0.05) between the products. As a result of these findings, snack sticks formulated with 0 and 3.50% SPC were compared in a consumer sensory evaluation. Consumer panelists detected no significant differences (P>0.05) in flavor, texture and overall acceptance between the snack stick products, and approximately 61% of the panelists commented that they would purchase them. Cabrito snack sticks formulated with 3.50% SPC had lower fat (P<0.05) and higher ash contents when compared with the control (0% SPC) sticks. Moisture, protein, water activity and smokehouse yields were similar (P>0.05) for both products. Moisture: protein ratio and pH values were higher (P<0.05) for snack sticks formulated with 3.5% SPC when compared with the control sticks. The addition of SPC resulted in a 4.60% reduction in the price of snack sticks formulated with 3.50% SPC when compared with control sticks.

Development and evaluation of a cabrito smoked sausage product.

In order for the meat goat producer to survive, new avenues for marketing goats must be created. Currently, the live animal is sold directly to consumers, or to brokers who in turn sell the animal directly to consumers or retail stores that cater to various ethnic groups. The production of value-added products with appeal to North American consumers, as well as current ethnic consumers, should result in increased profitability of the meat goat. The objectives of this study were to develop a value added product, cabrito smoked sausage, using goat meat as the sole meat ingredient; evaluate soy protein concentrate (SPC) at various levels in an effort to reduce product cost; determine consumer acceptability; and conduct a cost analysis to determine the approximate market price for the product. Three fermented cabrito smoked sausage products were manufactured containing 0, 1.75 or 3.50% SPC and stored at 2±1°C until evaluated. The sausages were evaluated for sensory characteristics, proximate analysis, pH, water activity and smokehouse yields. Trained panelists detected no significant flavor differences (P > 0.05) between the products. As a result of these findings, sausages formulated with 0 and 3.50% SPC were compared in a consumer sensory evaluation. Consumer panelists detected no significant differences (P > 0.05) in flavor, texture and overall acceptance between the snack sticks. Approximately 65% of the panelists commented that they would purchase the value added products. Proximates, pH, water activity and smokehouse yields were similar (P > 0.05) for the sausages formulated with 0 and 3.50% SPC. The addition of SPC resulted in an 8.79% reduction in the price of the 3.50% SPC formulation when compared to the sausage formulated with no SPC.