Chemical synthesis and structure elucidation of bovine K-casein (1-44)

The caseins (αs1, αs2, β, and κ) are phosphoproteins present in bovine milk that have been studied for over a century and whose structures remain obscure. Here we describe the chemical synthesis and structure elucidation of the N-terminal segment (1–44) of bovine κ-casein, the protein which maintains the micellar structure of the caseins. κ-Casein (1–44) was synthesised by highly optimised Boc solid-phase peptide chemistry and characterised by mass spectrometry. Structure elucidation was carried out by circular dichroism and nuclear magnetic resonance spectroscopy. CD analysis demonstrated that the segment was ill defined in aqueous medium but in 30% trifluoroethanol it exhibited considerable helical structure. Further, NMR analysis showed the presence of a helical segment containing 26 residues which extends from Pro8 to Arg34. This is the first report which demonstrates extensive secondary structure within the casein class of proteins.

Clinical signs and duration of cyanide toxicosis delivered by the M-44 ejector in wild dogs

Sodium cyanide poison is potentially a more humane method to control wild dogs than sodium fluoroacetate (1080) poison. This study quantified the clinical signs and duration of cyanide toxicosis delivered by the M-44 ejector. The device delivered a nominal 0.88 g of sodium cyanide, which caused the animal to loose the menace reflex in a mean of 43 s, and the animal was assumed to have undergone cerebral hypoxia after the last visible breath. The mean time to cerebral hypoxia was 156 s for a vertical pull and 434 s for a side pull. The difference was possibly because some cyanide may be lost in a side pull. There were three distinct phases of cyanide toxicosis: the initial phase was characterised by head shaking, panting and salivation; the immobilisation phase by incontinence, ataxia and loss of the righting reflex; and the cerebral hypoxia phase by a tetanic seizure. Clinical signs that were exhibited in more than one phase of cyanide toxicosis included retching, agonal breathing, vocalisation, vomiting, altered levels of ocular reflex, leg paddling, tonic muscular spasms, respiratory distress and muscle fasciculations of the muzzle.

Aiming for the management of the small hive beetle, Aethina tumida, using relative humidity and diatomaceous earth

Small hive beetles (SHBs) are a global pest of European honeybee colonies. In the laboratory, the survival of adult SHBs was evaluated in relation to relative humidity (RH = 56, 64, 73, 82 and 96 %) and treatment with diatomaceous earth (DE) across 4 days. Low RH reduced survival. The application of DE reduced survival in addition to RH. Adults treated with corn flour (control) showed no difference in survival from untreated beetles. Scanning electron microscopy images showed no scarification of adult beetle cuticle after exposure to DE; therefore, water loss is likely facilitated through non-abrasive means such as the adsorption of cuticular lipids. The data agree with the hypothesis that DE causes mortality through water loss from treated insects. Egress, ingress, mortality and the egg-laying behaviours of beetles were observed in relation to a popular in-hive trench trap with and without the addition of DE. Traps filled with DE resulted in 100 % mortality of beetles compared with 8.6 % mortality when no DE was present. A simple method for visually determining beetle sex was used and documented.

Welfare outcomes for 3- and 6-month-old beef calves in a tropical environment castrated surgically or by applying rubber rings

Castration of cattle using rubber rings is becoming increasingly popular due to the perceived ease of the procedure and greater operator safety when compared with surgical castration. Few comparative studies have investigated the effects of different castration methods and calf age on welfare outcomes, particularly in a tropical environment. Thirty Belmont Red (a tropically adapted breed), 3-month-old (liveweight 71–119 kg) and 30, 6-month-old (liveweight 141–189 kg) calves were assigned to a two age × three castration (surgical, ring and sham) treatment factorial study (Surg3, Surg6, Ring3, Ring6, Sham3 and Sham6, n = 10 for each treatment group). Welfare outcomes were assessed post-castration using: behaviour for 2 weeks; blood parameters (cortisol and haptoglobin concentrations) to 4 weeks; wound healing to 5 weeks; and liveweights to 6 weeks. More Surg calves struggled during castration compared with Sham and Ring (P < 0.05, 90 ± 7% vs. 20 ± 9% and 24 ± 10%) and performed more struggles (1.9 ± 0.2, 1.1 ± 0.3 and 1.1 ± 0.3 for Surg, Sham and Ring, respectively), suggesting that surgical castration caused most pain during performance of the procedure. A significant (P < 0.05) time × castration method × age interaction for plasma cortisol revealed that concentrations decreased most rapidly in Sham; the Ring6 calves failed to show reduced cortisol concentrations at 2 h post-castration, unlike other treatment groups. By 7 h post-castration, all treatment groups had similar concentrations. A significant (P < 0.01) interaction between time and castration method showed that haptoglobin concentrations increased slightly to 0.89 and 0.84 mg/mL for Surg and Ring, respectively over the first 3 days post-castration. Concentrations for Surg then decreased to levels similar to Sham by day 21 and, although concentrations for Ring decreased on day 7 to 0.76 mg/mL, they increased significantly on day 14 to 0.97 mg/mL before reducing to concentrations similar to the other groups (0.66 mg/mL) by day 21. Significantly (P < 0.05) more of the wounds of the 3-month compared with the 6-month calves scored as ‘healed’ at day 7 (74% vs. 39%), while more (P = 0.062) of the Surg than Ring scored as ‘healed’ at day 21 (60% vs. 29%). At day 14 there were significantly (P < 0.05) fewer healed wounds in Ring6 compared with other treatment groups (13% vs. 40–60%). Liveweight gain was significantly (P < 0.05) greater in 3-month (0.53 kg/day) than in 6-month calves (0.44 kg/day) and in Sham calves (P < 0.001, 0.54 kg/day), than in Ring (0.44 kg/day) and Surg (0.48 kg/day) calves. Overall, welfare outcomes were slightly better for Surg than Ring calves due to reduced inflammation and faster wound healing, with little difference between age groups.

Welfare outcomes for 3- and 6-month-old beef calves in a tropical environment castrated surgically or by applying rubber rings

Castration of cattle using rubber rings is becoming increasingly popular due to the perceived ease of the procedure and greater operator safety when compared with surgical castration. Few comparative studies have investigated the effects of different castration methods and calf age on welfare outcomes, particularly in a tropical environment. Thirty Belmont Red (a tropically adapted breed), 3-month-old (liveweight 71–119 kg) and 30, 6-month-old (liveweight 141–189 kg) calves were assigned to a two age × three castration (surgical, ring and sham) treatment factorial study (Surg3, Surg6, Ring3, Ring6, Sham3 and Sham6, n = 10 for each treatment group). Welfare outcomes were assessed post-castration using: behaviour for 2 weeks; blood parameters (cortisol and haptoglobin concentrations) to 4 weeks; wound healing to 5 weeks; and liveweights to 6 weeks. More Surg calves struggled during castration compared with Sham and Ring (P < 0.05, 90 ± 7% vs. 20 ± 9% and 24 ± 10%) and performed more struggles (1.9 ± 0.2, 1.1 ± 0.3 and 1.1 ± 0.3 for Surg, Sham and Ring, respectively), suggesting that surgical castration caused most pain during performance of the procedure. A significant (P < 0.05) time × castration method × age interaction for plasma cortisol revealed that concentrations decreased most rapidly in Sham; the Ring6 calves failed to show reduced cortisol concentrations at 2 h post-castration, unlike other treatment groups. By 7 h post-castration, all treatment groups had similar concentrations. A significant (P < 0.01) interaction between time and castration method showed that haptoglobin concentrations increased slightly to 0.89 and 0.84 mg/mL for Surg and Ring, respectively over the first 3 days post-castration. Concentrations for Surg then decreased to levels similar to Sham by day 21 and, although concentrations for Ring decreased on day 7 to 0.76 mg/mL, they increased significantly on day 14 to 0.97 mg/mL before reducing to concentrations similar to the other groups (0.66 mg/mL) by day 21. Significantly (P < 0.05) more of the wounds of the 3-month compared with the 6-month calves scored as ‘healed’ at day 7 (74% vs. 39%), while more (P = 0.062) of the Surg than Ring scored as ‘healed’ at day 21 (60% vs. 29%). At day 14 there were significantly (P < 0.05) fewer healed wounds in Ring6 compared with other treatment groups (13% vs. 40–60%). Liveweight gain was significantly (P < 0.05) greater in 3-month (0.53 kg/day) than in 6-month calves (0.44 kg/day) and in Sham calves (P < 0.001, 0.54 kg/day), than in Ring (0.44 kg/day) and Surg (0.48 kg/day) calves. Overall, welfare outcomes were slightly better for Surg than Ring calves due to reduced inflammation and faster wound healing, with little difference between age groups.