Experimental infection in calves with a specific subtype of verocytotoxin-producing Escherichia coli O157:H7 of bovine origin

Background

In Sweden, a particular subtype of verocytotoxin-producing Escherichia coli (VTEC) O157:H7, originally defined as being of phage type 4, and carrying two vtx₂ genes, has been found to cause the majority of reported human infections during the past 15 years, including both sporadic cases and outbreaks. One plausible explanation for this could be that this particular subtype is better adapted to colonise cattle, and thereby may be excreted in greater concentrations and for longer periods than other VTEC O157:H7 subtypes.

Methods

In an experimental study, 4 calves were inoculated with 10⁹ colony forming units (cfu) of strain CCUG 53931, representative of the subtype VTEC O157:H7 (PT4;vtx₂;vtx_2c). Two un-inoculated calves were co-housed with the inoculated calves. Initially, the VTEC O157:H7 strain had been isolated from a dairy herd with naturally occurring infection and the farm had previously also been linked to human infection with the same strain. Faecal samples were collected over up to a 2-month period and analysed for VTEC O157 by immuno-magnetic separation (IMS), and IMS positive samples were further analysed by direct plating to elucidate the shedding pattern. Samples were also collected from the pharynx.

Results

All inoculated calves proved culture-positive in faeces within 24 hours after inoculation and the un-inoculated calves similarly on days 1 and 3 post-inoculation. One calf was persistently culture-positive for 43 days; in the remainder, the VTEC O157:H7 count in faeces decreased over the first 2 weeks. All pharyngeal samples were culture-negative for VTEC O157:H7.

Conclusion

This study contributes with information concerning the dynamics of a specific subtype of VTEC O157:H7 colonisation in dairy calves. This subtype, VTEC O157:H7 (PT4;vtx_2;vtx_2c), is frequently isolated from Swedish cattle and has also been found to cause the majority of reported human infections in Sweden during the past 15 years. In most calves, inoculated with a representative strain of this specific subtype, the numbers of shed bacteria declined over the first two weeks. One calf could possibly be classified as a high-shedder, excreting high levels of the bacterium for a prolonged period.     

A preliminary study of Salmonella, verocytotoxigenic Escherichia coli/Escherichia coli O157 and Campylobacter on four mixed farms

The aims of this study were to investigate the incidence of Salmonella, verocytotoxigenic Escherichia coli (VTEC)/Escherichia coli O157 and Campylobacter on four mixed farms and to characterize the isolates in terms of a range of virulence factors. Eighty-nine composite (five different samples from the same animal species combined) faecal [cattle (24), pigs (14), sheep (4), poultry (4), horses (7), deer (4), dogs (9), rodents (2) and wild birds (20)] samples, 16 composite soil samples plus 35 individual water samples were screened using culture-based, immunomagnetic separation and molecular methods. Salmonella was detected in bovine faeces, cattle and poultry house water. Salmonella serotypes/phage types included Dublin, Kiel and Typhimurium DT193, and most isolates were spvC, invA and rck positive. The pefA and rck genes were found exclusively in the non-Typhimurium strains, while Salmonella Dublin and Salmonella Kiel strains carried Salmonella genomic island I marker(s). VTEC/E. coli O157 were found in deer and dog faeces only. The E. coli O157 isolate was an enteroinvasive E. coli, while the VTEC isolate was untypable but carried the vt1, eaeA, hlyA, tir and eptD genes. This article reports the first confirmed carriage of E. coli O157 in Irish deer. Campylobacter species were not detected over the course of this study. It was concluded that [1] Salmonella, VTEC and Campylobacter have low (<5%) prevalence or are absent on the farms in this study; [2] water was an important source of bacterial pathogens; [3] both dogs and deer may act as a source of pathogenic E. coli and [4] key virulence and resistance determinants are widespread in farm Salmonella strains. This study highlights the need to control water as a source of pathogens and suggests that the domestic pets and deer should be considered in any farm risk assessment.     

Salmonella Infantis and Salmonella Enteritidis specific bacteriophages isolated form poultry faeces as a complementary tool for cleaning and disinfection against Salmonella

Salmonellosis represents an important public health concern. Several authors point out the inefficiency of the cleaning and disinfection protocols to remove the bacteria from the field. For this reason, innovative techniques, as bacteriophages, could be implemented to control the bacteria. The main objectives of this study were to assess the effect of bacteriophages against Salmonella Infantis and Salmonella Enteritidis on farm surfaces, and to evaluate bacteriophage procedure application as sanitiser against Salmonella in field conditions. Thus, most prevalent serovars in poultry production were selected (Salmonella Infantis and Salmonella Enteritidis) to contaminate farm facilities. Then, two specific bacteriophages isolated from poultry faeces were applied against them. Results showed Salmonella Infantis and Salmonella Enteritidis decreased of 4.55 log₁₀CFU/mL and 3.85 log₁₀CFU/mL, respectively; the maximum reduction in Salmonella was the 5^th day, after 10⁸ PFU/mL and 10³ PFU/mL bacteriophage application. These results highlight bacteriophages as a promising tool together with cleaning and disinfection.     

In vivo activity of cefquinome against Riemerella anatipestifer using the pericarditis model in the duck

Cefquinome is a fourth‐generation cephalosporin with broad‐spectrum antibacterial activity, including activity against enteric gram‐negative bacilli such as Riemerella anatipestifer. The pericarditis model was used to examine the pharmacodynamic characteristics of cefquinome against R. anatipestifer. Serum levels of cefquinome following the administration of different doses were determined by LC‐MS/MS. Ducks with ca. 10⁶ CFU/mL at the initiation of therapy were treated with cefquinome at doses that ranged from 0.0156 to 2 mg/kg of body weight/day (in 3, 6, 12, or 24 divided doses) for 24 h. The percentage of a 24‐h dosing interval that the unbound serum cefquinome concentrations exceeded the MIC (fT > MIC) were the pharmacokinetic (PK)–pharmacodynamic (PD) parameter that best correlated with efficacy (R² 86.3% for R. anatipestifer, compared with 58.9% for the area under the concentration–time curve/MIC and 10.6% for peak/MIC). A sigmoid E_max model was used to estimate the magnitudes of the %fT > MIC associated with net bacterial stasis, a 1‐log₁₀ CFU reduction from baseline, and a 2‐log₁₀ CFU reduction from baseline; the corresponding values were (22.5 ± 1.3) %, (35.2 ± 4.5) %, and (42.4 ± 2.7) %. These data showed that treatment with cefquinome results in marked antibacterial effects in qvivo against R. anatipestifer and that the host's immunity may also play a key role in the anti‐infective therapy process.     

Lactobacillus plantarum BS22 promotes gut microbial homeostasis in broiler chickens exposed to aflatoxin B1

Probiotics promote the health of the host by maintaining intestinal microbial homeostasis. This study aimed to investigate the benefits of Lactobacillus plantarum BS22 (LP) in the gastrointestinal tract (GIT) microbial homeostasis of broiler chickens exposed to aflatoxin B₁ using the PCR‐DGGE, viable count and real‐time PCR. The toxin adsorption experiment demonstrated that treatment R5 (1.0 × 10⁸ CFU/g LP) exhibited good absorptive effect in adsorbing the aflatoxin B₁ (AFB₁) in vitro. DGGE showed that the composition and structure of gut microbiota were more similar in the mucosa than in the content of all the samples. In addition, higher diversity of the microbiota was observed in the caecum and glandular stomach than in other segments. Lactobacillus, Enterococcus and Enterobacteriaceae were more abundant in the ileum than in the other segments. Enterobacteriaceae in groups I (basal diet) and II (basal diet+50 μg/kg AFB₁) showed a significant difference in group III (basal diet + 50 μg/kg AFB₁ + 1 × 10⁸ CFU/g LP) in the crop content and duodenum mucosa (p < .05). This investigation indicates that the L. plantarum BS22 promotes GIT microbial homeostasis in broiler chickens exposed to AFB₁, particularly for the intestine mucosa microbiota. Thus, L. plantarum BS22 is a possible candidate for degrading AFB_1.     

Permeability of the endothelium and partitioning of the pulmonary blood flow resistance in isolated perfused pig lungs: Effects of breed and age

The right and left lungs of 5 healthy Minipigs and of 13 healthy Landrace piglets were isolated, perfused at constant pressure and maintained in an isogravimetric state under zone III conditions (pulmonary venous pressure>alveolar pressure). By applying the double, arterial and venous, occlusion technique, the total blood flow resistance (R _t) was partitioned into four components: arterial (R _a), pre-(R _a) and post-capillary (R _v) and venous (R _v). The capillary filtration coefficient (K _f,c) was evaluated by measuring the weight gained by the lungs when the arterial and venous pressures were suddenly increased. In the youngest Landrace piglets (5 weeks old), there was an uncontrolled vasoconstriction which sometimes prevented perfusion of the lungs and induced a large increase inR _t. These high values ofR _t were decreased by tolazoline administration. The values ofR _t recorded in older pigs (12–13 weeks old) were lower in Minipigs (33.66±3.77 cmH₂O min L^–1 per 100 g of lungs;n=5) than in Landrace piglets (55.20±6.18 cmH₂O min L^–1 per 100 g;n=5). This breed difference was due to the differences inR _a andR _v. The mean values ofK _f,c were 0.193±0.015 and 0.202±0.029 ml min (cmH₂O)^–1 per 100 g of the lungs in Minipigs and Landrace piglets respectively. All these parameters were stable for the 3 hours following the equilibrium period. It was concluded that: (1) There is an age-related maturation of the control of the vasomotor tone in porcine lungs. (2) Pulmonary microvascular haemodynamics are influenced by the breed of the pigs. (3) There was no difference in theK _f,c values between both the breeds. (4) A comparison of the values reported for dogs and rabbits with our data shows that the pre- and post-capillary resistances and, to a lesser extent, the arterial and venous resistances are relatively high in pigs.     

Verocytotoxin-producing Escherichia coli O157 on a farm open to the public: outbreak investigation and longitudinal bacteriological study

Verocytotoxin-producing Escherichia coli (VTEC) O157 phage type 2 (PT2) was isolated from three calves and two goats on a farm open to the public. Phenotypic and DNA-based typing showed that the strains were identical or very closely related to those obtained from an outbreak of VTEC O157 infection in two separate family groups who visited the farm. No VTEC O157 PT2 was isolated again from the farm during a 12-month longitudinal bacteriological study undertaken after the infected animals had been removed. However, phenotypically and genotypically indistinguishable VTEC O157 PT2/28 strains were detected in two of 474 faecal samples collected at monthly visits from 15 species of animals of various ages. The two isolates were obtained from calves from different sources sampled 146 days apart, suggesting that the infection had persisted on the farm although it was not detected in the other species. The same strain was subsequently isolated from another calf housed in the same pen as one of the infected calves. The longest period during which the organism was excreted was seven days. No VTEC O157 was isolated either from 204 replacement animals (including 138 orphan lambs and 10 calves) brought in from various sources, and sampled while they were kept in isolation for two weeks before being introduced to the farm, or from environmental samples. During the study a visitor became ill with VTEC O157 PT2. However, the isolate was distinct from those recovered from the farm and there was no evidence to suggest that the visit was the source of the infection.     

Efficacy assessment of an intramammary treatment with a new recrystallized enrofloxacin vs ceftiofur and parenteral enrofloxacin in dairy cows with nonsevere clinical mastitis

A recrystallized form of enrofloxacin as dehydrate‐HCl (enro‐C) was assessed for bacteriological and clinical cure efficacies in Holstein‐Friesian cows affected of nonsevere clinical mastitis. Treatments were enro‐C_susp (n = 81), treated with a pharmaceutical suspension of enro‐C/quarter; group enro‐C_pd (n = 80) treated as above, but using enro‐C powder suspended in water; group CF (n = 65), treated with ceftiofur HCl/quarter; and group enro_R (n = 66), treated with standard enrofloxacin solution (5 mg/kg, intramuscular). Cows had a mean milk production of 31 L/day and were 2‐3 lactational periods old. Treatments were administered every 24 hr for 3 days. Groups treated with enro‐C exhibited statistically significant (p > .05) better clinical cure as compared to groups treated with CF or enro_R (95.06%, 96.25%, 67.79%, and 57.55%, for enro‐C_susp, enro‐C_pd, CF, and enro_R, respectively). In contrast, probability of bacteriological cure was not statistically different among treatments. Yet, the outstanding clinical and bacteriological cure rates obtained for enro‐C for nonsevere cases of mastitis is superior to previously reported data for parenteral enrofloxacin and other antibacterial‐intramammary treatments. Impact of using enro‐C on the rate and pattern of bacterial resistance, somatic cell counts and milk electric conductivity, must be studied. Also, the use of enro‐C for complicated cases of mastitis should be studied and milk withdrawal times must be accurately established.     

Age- and sex-dependent regression models for predicting the live weight of West African Dwarf goat from body measurements

The relationships between live weight and eight body measurements of West African Dwarf (WAD) goats were studied using 211 animals under farm condition. The animals were categorized based on age and sex. Data obtained on height at withers (HW), heart girth (HG), body length (BL), head length (HL), and length of hindquarter (LHQ) were fitted into simple linear, allometric, and multiple-regression models to predict live weight from the body measurements according to age group and sex. Results showed that live weight, HG, BL, LHQ, HL, and HW increased with the age of the animals. In multiple-regression model, HG and HL best fit the model for goat kids; HG, HW, and HL for goat aged 13–24 months; while HG, LHQ, HW, and HL best fit the model for goats aged 25–36 months. Coefficients of determination (R ²) values for linear and allometric models for predicting the live weight of WAD goat increased with age in all the body measurements, with HG being the most satisfactory single measurement in predicting the live weight of WAD goat. Sex had significant influence on the model with R ² values consistently higher in females except the models for LHQ and HW.     

Intake,digestibility and nitrogen balance in Mithun (<Emphasis Type="BoldItalic">Bos frontalis</Emphasis>) offered urea-treated paddy straw based feed blocks

The aim of the study was to determine the effect of feeding feed blocks containing varying proportion of urea-treated paddy straw (UTPS) on dry matter (DM) intake (DMI), nutrient utilization and N balance in Mithun. For the purpose, four adult male Mithun (279.5 ± 8.2 kg) were selected and offered four experimental rations viz. R₁ (Napier fodder + concentrate at 60:40), R₂ (UTPS + concentrate at 50:50), R₃ (UTPS + concentrate at 60:40) and R₄ (UTPS + concentrate at 70:30) in 4 × 4 Latin square design. The DMI % of body weight was 2.59, 2.96, 2.85 and 2.77 and the DMI g kg⁻¹ W^0.75 was 107, 123, 118 and 115 in Mithun fed R₁, R₂, R₃ and R₄, respectively. The mean DMI was (P < 0.01) higher in animals fed R₂ and R₃ than R₁ and R₄, whereas the water intake was (P < 0.01) higher in Mithun fed R₂, R₃ and R₄ than R₁. The digestibility of DM, organic matter, crude protein, crude fibre, neutral detergent fibre and cellulose were (P < 0.05) higher in animals fed R₂, R₃ and R₄ than R₁. A positive N balance was observed in all the experimental animals, with higher (P < 0.05) values among the animals offered R₂, R₃ and R₄ than R₁. The digestible crude protein and total digestible nutrient intakes were higher (P < 0.05) in Mithun fed R₂ and R₃ than R₁ and R₄. It is concluded that the UTPS can be incorporated up to 70% to formulate the complete feed/feed block and can be used for feeding of Mithun under complete confinement system.     

Inheritance of the lace‐tailed laced plumage pattern of the sebright bantam

1. An investigation was conducted firstly among progeny from a cross between Blue Andalusian and Silver‐spangled Hamburgh bantams, and secondly between crosses of selected progeny of this mating with a Silver Sebright bantam in an effort to determine the genotype of the lace‐tailed laced plumage phenotype of the latter.

2. The genotype of the black‐laced blue and the spangled plumage phenotypes of the Andalusian and the Silver‐spangled Hamburgh had, respectively, been shown to depend on homozygosity of E, Co, db⁺, Ml and Pg, and of co⁺, Db, Ml and Pg together with a black down allele at the E‐locus presumed to be E, but also hypothesised to be E^R. The genes E and E^R are the extended black and birchen‐like allele at the E‐locus whilst Co, Db, Ml and Pg are, respectively, the eumelanin restrictors, Columbian and dark‐brown Columbian, the eumelanin extension melanotic and the pattern gene. The Sebright had been hypothesised to possess the E allele at the E‐locus, and to be homozy‐gous Co, Ml and Pg, a combination shown to be responsible for the black‐tailed laced phenotype of the Wyandotte.

3. Segregation in the F₂ generation varied from that expected if both parental genotypes were E/E, but gave close agreement if the Silver‐spangled Hamburgh was E^R/E^R.

4. A lace‐tailed laced segregant in the F₂ generation of the first mating, presumed to be homozygous E^R, Co, Db, Ml and Pg was mated to a Sebright. The F₁ generation failed to segregate at the 5 loci, thus suggesting the genotype of the lace‐tailed laced phenotype of the Sebright to be homozygous E^R, Co, Db, Ml and Pg. Segregation in the F₂ generation of a mating of the female F₁ with a Silver‐spangled Hamburgh male confirmed the genotype of the lace‐tailed lacing of the Sebright bantam, and demonstrated that of the Silver‐spangled Hamburgh to be homozygous E^R, co⁺, Db, Ml and Pg‐.     

Verocytotoxin-producing Escherichia coli (VTEC)

Escherichia coli O157:H7 and other Verocytotoxin-producing E. coli (VTEC) are zoonotic pathogens associated with food and waterborne illness around the world. E. coli O157:H7 has been implicated in large outbreaks as well as in sporadic cases of haemorrhagic colitis and the sometimes fatal haemolytic uremic syndrome. VTs produced by these bacteria are thought to damage host endothelial cells in small vessels of the intestine, kidney and brain resulting in thrombotic microangiopathy. All VTs have the same subunit structure, glycolipid cell receptor and inhibit protein synthesis. During VTEC infection, it is thought one or more bacterial adhesins initiates colonization and establishes intimate attachment and is responsible for the translocation of a variety of effectors which alter the structure and function of host cells. VTEC are widespread in animals but ruminants are thought to be their natural reservoir. E. coli O157:H7 colonizes the terminal colon of cattle and can be shed in very large numbers by specific herdmates known as “supershedders”. Faeces containing these organisms act as a source of contamination for a variety of foods and the environment. Many VTEC control efforts have been investigated along the “farm to fork” continuum including, vaccination of cattle with colonization factors, and the use of novel antimicrobials, such as bacteriocins, chloral hydrate, bacteriophage and substances which disrupt quorum sensing. In addition, many barriers have been developed for use in the slaughter and food processing industry such as steam pasteurization and irradiation. Despite these efforts many scientific, technical and regulatory challenges remain in the control and prevention of VTEC-associated human illness.     

Comparative Study of Nitrogen Metabolism and the Nitrogen Maintenance Requirement in Lohmann Brown Roosters and Layers

The objective of this study was to investigate the effect of dietary protein level on nitrogen retention and compare nitrogen metabolism in Lohmann Brown adult roosters and layers. The nitrogen maintenance requirement (N_m) was determined based on the nitrogen balance. Thirty Lohmann Brown adult roosters and 30 laying hens at 27 weeks of age were randomly divided into five groups of six birds per group. The birds were fed with one of five diets containing 10.46%, 11.77%, 13.79%, 16.77% or 18.29% of crude protein. Nitrogen intake, nitrogen retention and nitrogen retention efficiency were higher in roosters than in laying hens, and the average nitrogen retention rate for groups fed with CP level of 11.77%, 13.79%, 16.77% and 18.29% was improved by 9.14%. The nitrogen maintenance requirement for Lohmann Brown roosters and laying hens at 27 weeks of age were 0.4245 g/d and 0.5059 g/d, respectively, and Nm based on average body weight (BW) and metabolic body weight (BW^0.75) was 0.2364 g/kg BW and 0.2739 g/kg BW^0.75 for laying hens and 0.2754 g/kg BW and 0.3208 g/kg BW^0.75 for roosters, respectively. The regression equations for daily N gain (N_B, protein accretion) vs. daily N intake (N_I) for Lohmann Brown layers and roosters were N_B = 0.3743N_I–0.1589 (R² = 0.79) and N_B = 0.6228N_I–0.3151 (R² = 0.85), respectively. The results of this study indicate that nitrogen intake and nitrogen retention at the same dietary CP level were higher in roosters than in laying hens.     

Physicochemical characterization and pharmacokinetics in broiler chickens of a new recrystallized enrofloxacin hydrochloride dihydrate

Enrofloxacin, a key antimicrobial agent in commercial avian medicine, has limited bioavailability (60%). This prompted its chemical manipulation to yield a new solvate‐recrystallized enrofloxacin hydrochloride dihydrate entity (enro_C). Its chemical structure was characterized by means of mass spectroscopy, Fourier transformed infrared spectroscopy, X‐ray powder diffraction, and thermal analysis. Comparative oral pharmacokinetics (PK) of reference enrofloxacin (enro_R) and enro_C in broiler chickens after oral administration revealed noticeable improvements in key parameters and PK/PD ratios. Maximum serum concentration values were 2.61 ± 0.21 and 5.9 ± 0.42 μg/mL for enro_R and enro_C, respectively; mean residence time was increased from 5.50 ± 0.26 h to 6.20 ± 0.71 h and the relative bioavailability of enro_C was 336%. Considering C_max/MIC and AUC/MIC ratios and the MIC values for a wild‐type Escherichia coli O78/H12 (0.25 μg/mL), optimal ratios will only be achieved by enro_C (C_max/MIC = 23.6 and AUC/MIC = 197.7 for enro_C; vs. C_max/MIC = 10.4 and AUC/MIC = 78.1 for enro_R). Furthermore, enro_C may provide in most cases mutant prevention concentrations (C_max/MIC ≥ 16). Ready solubility of powder enro_C in drinking water at concentrations regularly used (0.01%) to provide an additional advantage of enro_C in the field. Further development of enro_C is warranted before it can be recommended for clinical use in veterinary medicine.     

Endotoxin-induced microvascular injury in isolated and perfused pig lungs

The lungs of 13 healthy Landrace piglets were isolated, perfused and maintained in an isogravimetric state under zone III conditions. By applying vascular occlusion methods, the total blood flow resistance (Rt) was partitioned into four components: arterial (Ra), pre- (Ra) and post-capillary (Rv), and venous (Rv). The capillary filtration coefficient (K _f,c) was evaluated using a gravimetric technique. A bolus of 55 µg ofEscherichia coli endotoxins (LPS) per 100 g of lung was injected into the arterial reservoir of eight lungs, followed by an infusion of LPS at a rate of 55 µg per 100 g of lung per hour for 180 min. A bolus of theophylline (85 mg per 100 g of lung weight) was injected into the arterial reservoir after the last determination of theK _f,c value. All the parameters were evaluated again when the lungs reached a new steady state. Endotoxin induced a significant increase inRt from 54.7 ± 7.0 at zero time to 184.7±44.2 cm H₂O min L^–1 (100 g)^–1 180 minutes later, which can be ascribed to the increase inRa andRv. These haemodynamic modifications were related to the increases in the arterial pressure and in the pressure at the distal end of the arterial segment and to the decreases in the pressure at the proximal end of the venous segment and in the blood flow. The capillary pressure and the lung weight remained unchanged. Endotoxin infusion infusion induced an increase in theK _f,c value from 0.208±0.011 (att=0) to 0.391±0.034 ml min^–1 (cmH₂O)^–1 (100 g)^–1 (att=180). Administration of theophylline significantly reducedRt,Ra,Ra andRv towards or under the baseline values and also induced a significant increase in the lung weight and in theK _f,c value. It was concluded that the endotoxin-induced increase in the total blood flow resistance can be ascribed to a vasospasm occurring at the level of the pre- and post-capillary small vessels and that changes in the permeability of the endothelium greatly contribute to the development of the pulmonary oedema observed in endotoxaemic pigs.     

Three generations of selection for high and low donor competences of splenomegaly in chickens

Three generations of two‐way selection were conducted for the donor competence of graft‐versus‐host reaction. Selection was effective in changing the donor competence. The average realised heritability over three generations was determined to be 0.41 by measuring the difference between the high (H) and low (L) lines. Reciprocal crosses gave intermediate values compared with those for their respective line offspring.

Changes of gene frequencies at the B blood group locus were examined. In the H line the frequency of B ¹¹ allele increased and of B ⁹ decreased, as expected, after selection in each generation. In the L line, however, the expected change was observed only in the first two generations. The direction of change reversed in the latter two generations. Relationship between the B locus genotypes and the degree of splenomegaly was different according to the sex of donors. In male donors, the B ⁹ /B ¹¹ induced larger spleen than the B ⁹ /B ⁹ did, but in females the B ⁹ /B ⁹ was generally larger than the B ⁹ /B ¹¹ especially in the L line.     

Effect of supplementation with MgCO3 and l-Tryptophan on the welfare and on the carcass and meat quality of two halothane pig genotypes (NN and nn)

Sixty-one animals with different Halothane genes (homozygous halothane positive, n = 34; and homozygous halothane negative, n = 27) were fed with three diets (control group, with no supplement; magnesium (Mg) group with 1.28 g MgCO₃/kg and tryptophan (Trp) group with 5 g l-Trp/kg) during the last 5 days before slaughter. Animals were submitted to minimal stress antemortem conditions. Pig behaviour was recorded at the experimental farm, raceway to the CO₂ stunning system and during the stunning period. Corneal reflexes were recorded after stunning as well. There were no differences in feed intake among diets (P > 0.05) during the 5 days of treatment. The halothane positive (nn) group had lower intake than the halothane negative (NN) group (P < 0.01). The behaviour of the pigs in the raceway did not differ (P > 0.05) among treatments or halothane genotype. A significant (P < 0.001) interaction diet  halothane was found in the time to appear the first retreat attempt during the exposure to the CO₂ system. In the nn group, the time of performing the first retreat attempt was later in the Mg (P < 0.05) than the control group. Moreover, in the Mg group, the nn had a later (P < 0.05) first retreat attempt than the NN. Thus, Mg supplementation could have a positive effect on welfare of nn pigs. The nn had a lower proportion of animals that showed corneal reflexes after stunning than NN, indicating a higher effectiveness of the stunning method in nn pigs. Neither Mg nor Trp affected carcass quality and meat quality parameters, although significant differences were found between genotypes.     

Genetic structure of milk protein polymorphisms and effects on milk production traits in a local dairy cattle

The milk protein genetic structure of the Reggiana dairy local cattle with remarkable milk quality properties was analysed and compared with the structure of Italian Friesian and Italian Brown cattle. A total of 1138 individual milk samples from the three breeds were typed by isoelectrofocusing. Polymorphism was found at the α_s1‐casein (CSN1S1), β‐casein (CSN2), κ‐casein (CSN3), and β‐lactoglobulin (LGB) loci. High frequencies were observed for some casein alleles in Reggiana (CSN1S1*C, 25%; CSN2*B: 28%) with considerable differences to those observed in a survey carried out 30 years before. A great variation among breeds was observed for casein haplotype frequencies and high linkage disequilibrium was detected at the three polymorphic casein loci CSN1S1‐CSN2‐CSN3. The prevalent haplotypes were CA²B (23%; Reggiana), BA²A (48%; Italian Friesian) and BA²B (51%; Italian Brown). Higher G_ST values between breeds were observed when haplotype frequencies were considered instead of gene frequencies. In the Reggiana cattle, the occurrence of two casein alleles at a rather high frequency (CSN1S1*C, CSN2*B) allowed to estimate the effects of rare casein haplotypes on milk traits. The CA²B haplotype was associated with a significantly higher milk protein and fat content and a significantly higher protein per cent was found also for the BA¹B haplotype.