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1.
Poult Sci ; 103(6): 103698, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38657523

ABSTRACT

This study aimed to compare the inclusion of transgenic sorghums against commercially available sorghums on growth performance in broiler chickens. Isonitrogenous and isoenergetic diets were offered to a total 288 male Ross 308 broiler chickens from 14 to 35 d posthatch. Three dietary treatments were diets based on transgenic sorghums with a mean protein content of 154.7 g/kg and 5 treatments were based on commercially available sorghum hybrids with a mean protein content of 90.6 g/kg. Soybean meal inclusions in the commercial sorghum diets averaged 215 g/kg, which was reduced to 171 g/kg in the transgenic sorghum diets because of the higher protein contents. Overall growth performance was highly satisfactory, and commercial sorghums supported 2.55% (2,330 vs. 2,272 g/bird; P = 0.010) more weight gains and 2.74% (2,929 vs. 2,851 g/bird; P = 0.012) higher feed intakes; however, the transgenic sorghums supported a fractionally better FCR (1.255 vs 1.257; P = 0.826). There were no statistical differences in apparent jejunal and ileal starch and protein (N) digestibility coefficients between treatments. The transgenic sorghum diets generated slightly, but significantly, higher AME:GE ratios and AMEn, but the commercial sorghum diets generated 6.33% (235 vs. 221 g/kg; P < 0.001) greater breast meat yields. Apparent ileal digestibility coefficients of 16 amino acids averaged 0.839 and 0.832 for transgenic and commercial sorghum-based diets, respectively, without any significant differences in individual amino acids. This outcome suggests amino acid digestibilities of the transgenic sorghums may be inherently higher than commercial hybrid sorghums as the 25.7% higher average soybean meal inclusions would have advantaged amino acid digestibilities in commercial sorghum diets. The possibility that the digestibilities of amino acids in the kafirin component of transgenic sorghums was enhanced by modifications to the structure of kafirin protein bodies is discussed. In conclusion, transgenic sorghums with higher protein concentrations led to 20.5% reduction of soybean meal inclusions in broiler diets, and this change did not compromise feed conversion efficiency compared to standard commercial hybrid sorghums.


Subject(s)
Animal Feed , Animal Nutritional Physiological Phenomena , Chickens , Diet , Plants, Genetically Modified , Sorghum , Animals , Sorghum/chemistry , Animal Feed/analysis , Male , Diet/veterinary , Chickens/physiology , Chickens/growth & development , Chickens/genetics , Digestion , Random Allocation , Dietary Proteins/metabolism , Diet, High-Protein/veterinary
2.
Res Vet Sci ; 169: 105158, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38295629

ABSTRACT

"Exploring AMH Levels, Homeostasis and Primordial Follicle Activation in Pubertal Infected Sheep on a High Protein Diet ". The first activation wave of ovarian primordial follicles is part of the onset of puberty and fertility. Abomasal helminth infection may cause an undesirable delay in puberty manifestation. Helminth-infected animals demand a higher amount of protein in their diet to repair the damage caused by the parasite in sheep's tissues, replenish the blood losses, and build the host's immune response. Helminths become resistant to drug therapy shortly after being exposed to a new treatment. Besides, there is the possibility of contamination by anthelmintic drugs in ovine products, possibly affecting human health and the environment. This study's objective was to evaluate if ovarian and clinical parameters can be improved by supplementing their diet with protein, offering a more sustainable management approach than relying on anthelmintic usage. We used a 2 × 2 factorial model where eighteen ewe lambs (Ovis aries) between 6 and 7 months old - born to the same ram - were fed one of two diet protein levels (12% or 19%). After 35 days on this diet, they were infected or left uninfected with 10,000 Haemonchus contortus L3 larvae. We evaluated Anti-Mullerian Hormone serum levels, blood cells and biochemical parameters at four different time points. Following 42 days of infection and 77 days on the diet, the lambs had their left ovaries removed, and we examined ovarian morphometrics through histological analysis. The groups Supplemented Protein-Infected(n = 5), Control Protein- Infected(n = 5), Supplemented Protein-Not Infected (n = 4) and Control Protein-Not Infected (n = 4) did not differ in their bodyweight gain. In the factorial ANOVA analysis examining the relationship between plasma protein, diet, and infection, the protein level of the diet showed significance (p = 0.02). Primordial follicle size varied with the interaction between diet and infection (p < 0.05), and oocyte size was affected by the level of protein in the diet (p = 0.047). Additionally, to understand how all homeostasis parameters relate to the primordial follicle and oocyte size, we applied an explanatory linear mixed model. In conclusion, serum AMH levels remained stable despite the infection and variations in diet protein levels, indicating its reliability as a marker for ovarian reserve in pubertal sheep. The number of blood cells, biochemical parameters, and primordial follicle activation were affected by both diet and infection.


Subject(s)
Anthelmintics , Diet, High-Protein , Female , Animals , Sheep , Male , Humans , Ovary , Anti-Mullerian Hormone/metabolism , Reproducibility of Results , Diet, High-Protein/veterinary
3.
J Dairy Sci ; 106(6): 4002-4017, 2023 Jun.
Article in English | MEDLINE | ID: mdl-37105871

ABSTRACT

We previously observed that diets with reduced starch concentration decreased yields of milk and milk protein in dairy cows fed low metabolizable protein diets. Supplementation of reduced-starch diets with a lipid source may attenuate or eliminate production losses. Our objective was to investigate the effects of partially replacing ground corn with soyhulls plus a palmitic acid-enriched supplement on dry matter (DM) intake, milk yield and composition, plasma AA concentration, and N and energy utilization in cows fed low metabolizable protein diets (mean = -68 g/d balance) with or without rumen-protected Met, Lys, and His (RP-MLH). Sixteen multiparous Holstein cows averaging (mean ± standard deviation) 112 ± 28 d in milk, 724 ± 44 kg of body weight, and 46 ± 5 kg/d of milk in the beginning of the study were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Each period lasted 21 d, consisting of 14 d for diet adaptation and 7 d for data and sample collection. Diets were fed as follows: (1) high starch (HS), (2) HS plus RP-MLH (HS+AA), (3) reduced starch plus a palmitic acid-enriched supplement (RSPA), and (4) RSPA plus RP-MLH (RSPA+AA). The HS diet contained (DM basis) 26% ground corn and 7% soyhulls, and the RSPA diet had 10% ground corn, 22% soyhulls, and 1.5% palmitic acid. The HS diet averaged (DM basis) 32.6% starch and 4% ether extract, while starch and ether extract concentrations of the RSPA diet were 21.7 and 5.9%, respectively. All 4 diets had (DM basis) 40% corn silage, 5% mixed-mostly grass haylage, 5% grass hay, and 50% concentrate. Diets did not affect DM intake and milk yield. Contrarily, feeding RSPA and RSPA+AA increased yields of energy-corrected milk (47.0 vs. 44.8 kg/d) and milk fat (1.65 vs. 1.50 kg/d) compared with HS and HS+AA. Milk fat concentration tended to decrease when RP-MLH was supplemented to HS, but no change was seen when added to RS (starch level × RP-MLH interaction). Milk and plasma urea N increased, and milk N efficiency decreased in cows fed RSPA and RSPA+AA versus HS and HS+AA. Apparent total-tract digestibilites of crude protein and neutral detergent fiber, as well as urinary urea N and total N excretion, were greater in cows offered RSPA and RSPA+AA than HS and HS+AA. Plasma Met and His concentrations increased with supplemental RP-MLH. Intake of gross energy and digestible energy and the output of urinary and milk energy were all greater with feeding RSPA and RSPA+AA versus HS and HS+AA. In summary, partially replacing ground corn with soyhulls plus palmitic acid in diets supplemented or not with RP-MLH increased milk fat yield and fiber digestibility and maintained DM intake and milk yield, but with decreased milk N efficiency and elevated urinary N excretion.


Subject(s)
Animal Feed , Diet, High-Protein , Glycine max , Palmitic Acid , Zea mays , Diet, High-Protein/veterinary , Amino Acids , Rumen , Animals , Cattle , Starch , Dietary Supplements , Nutrients
4.
J Anim Sci ; 1012023 Jan 03.
Article in English | MEDLINE | ID: mdl-36807528

ABSTRACT

Given the dynamic market for protein-based ingredients in the pet food industry, demand continues to increase for both plant- and animal-based options. Protein sources contain different amino acid (AA) profiles and vary in digestibility, affecting protein quality. The objective of this study was to evaluate the apparent total tract digestibility (ATTD) of canine diets differing in protein source and test their effects on serum metabolites and fecal characteristics, metabolites, and microbiota of healthy adult dogs consuming them. Four extruded diets were formulated to be isonitrogenous and meet the nutrient needs for adult dogs at maintenance, with the primary difference being protein source: 1) fresh deboned, dried, and spray-dried chicken (DC), 2) chicken by-product meal (CBPM), 3) wheat gluten meal (WGM), and 4) corn gluten meal (CGM). Twelve adult spayed female beagles (body weight [BW] = 9.9 ± 1.0 kg; age = 6.3 ± 1.1 yr) were used in a replicated 4 × 4 Latin square design (n = 12/treatment). Each period consisted of a 22-d adaptation phase, 5 d for fecal collection, and 1 d for blood collection. Fecal microbiota data were analyzed using QIIME 2.2020.8. All other data were analyzed using the Mixed Models procedure of SAS version 9.4. Fecal scores were higher (P < 0.05; looser stools) in dogs fed DC or CBPM than those fed WGM or CGM, but all remained within an appropriate range. Dry matter ATTD was lower (P < 0.05) in dogs fed CBPM or CGM than those fed DC or WGM. Crude protein ATTD was lower (P < 0.05) in dogs fed DC or CGM than those fed WGM. Dogs fed CBPM had lower (P < 0.05) organic matter, crude protein, and energy ATTD than those fed the other diets. Fecal indole was higher (P < 0.05) in dogs fed CBPM than those fed WGM. Fecal short-chain fatty acids were higher (P < 0.05) in dogs fed DC than those fed CGM. Fecal branched-chain fatty acids were higher (P < 0.05) in dogs fed DC or CBPM than those fed WGM. Fecal ammonia was higher (P < 0.05) in dogs fed DC or CBPM than those fed WGM or CGM. The relative abundances of three bacterial phyla and nine bacterial genera were shifted among treatment groups (P < 0.05). Considering AA profiles and digestibility data, the DC diet protein sources provided the highest quality protein without additional AA supplementation, but the animal-based protein diets resulted in higher fecal proteolytic metabolites. Further studies evaluating moderate dietary protein concentrations are needed to better compare plant- and animal-based protein sources.


Pet food trends are constantly changing. Because consumers are often focused on dietary proteins, with ingredient sources, dietary inclusion levels, and processing methods being important, they are a popular research topic. Protein sources contain different amino acid (AA) profiles and vary in digestibility, affecting protein quality. Our objective was to evaluate the apparent total tract digestibility of canine diets differing in protein source and test their effects on serum metabolites and fecal characteristics, metabolites, and microbiota of healthy adult dogs. Test diets were formulated to be similar nutritionally, but differed in protein source: fresh deboned, dried, and spray-dried chicken (DC), chicken by-product meal (CBPM), wheat gluten meal (WGM), and corn gluten meal (CGM). Fecal scores were higher in dogs fed chicken-based diets, but remained within an appropriate range. Dogs fed CBPM had lower nutrient and energy digestibilities than those fed the other diets, with protein digestibility also being lower in dogs fed DC or CGM than those fed WGM. Fecal metabolites and microbiota were shifted among diets, with animal-based protein diets increasing fecal protein metabolites. All diets were complete and balanced and performed well. When considering AA profiles and digestibility, however, the DC diet provided the highest protein quality.


Subject(s)
Diet, High-Protein , Digestion , Dogs , Animals , Feces/chemistry , Diet/veterinary , Diet, High-Protein/veterinary , Amino Acids/metabolism , Glutens/analysis , Animal Feed/analysis , Animal Nutritional Physiological Phenomena
5.
Poult Sci ; 101(4): 101724, 2022 Apr.
Article in English | MEDLINE | ID: mdl-35196586

ABSTRACT

Reflux of urine from the cloaca into the ceca provides chickens with a mechanism for recycling of urinary-Nitrogen (N) in a way analogous to urea recycling in mammals. However, it is unknown if reflux has substantial relevance in current poultry husbandry, where birds are fed ad libitum and have high protein intake. To evaluate the fate of urinary-N in ad libitum-fed broiler chickens, 15-day-old broilers were assigned to a high (21.9% CP, n = 22) or low (10.2 % CP, n = 22) protein diet. At 25 d of age, 20 broilers per dietary treatment were infused into the cloaca with a pulse dose of 107 mg [1,3-15N]-uric acid. N-contents and 15N-enrichment in digesta, blood plasma, and body tissues were measured at 5, 30, 60, 90, 150, 300, 450, 600, 1,200, or 1,800 min after administration (n = 2 /time-point /diet). Two broilers per dietary treatment were infused with saline and served as control to analyze background 15N-enrichment. The average total recovery (% of infused (w/w)) of 15N from infused uric acid in all body tissues was low (2.9 ± 0.62 %), of which the largest proportion was found in carcass tissue (2.5 ± 0.60%). 15N-enrichment was greatest in intestinal tissues. Even at 1,200 min, 15N-enrichment of ceca (0.46 ± 0.169 APE) and colon (0.13 ± 0.159 APE) digesta was considerably exceeding background enrichment. 15N-enrichment in excess of background enrichment in cecum and colon digesta (10-fold, P < 0.05), and 15N recovery in intestinal tissues (4-fold, P < 0.01) were greater in birds fed the low protein diet compared with the high protein diet, speculatively pointing out differences in the occurrence of reflux, incorporation of uric acid-N derivatives in intestinal tissues by first-pass metabolism, and a prolonged digesta retention time in protein deficient birds. In conclusion, these data confirm that uric acid-N infused in the cloaca can be refluxed and used for body N-deposition, but its contribution to whole body protein metabolism in broilers is probably limited.


Subject(s)
Chickens , Diet, High-Protein , Animal Feed/analysis , Animal Nutritional Physiological Phenomena , Animals , Chickens/metabolism , Diet/veterinary , Diet, High-Protein/veterinary , Diet, Protein-Restricted/veterinary , Dietary Supplements , Mammals , Uric Acid/metabolism
6.
J Anim Physiol Anim Nutr (Berl) ; 106(6): 1277-1287, 2022 Nov.
Article in English | MEDLINE | ID: mdl-34719816

ABSTRACT

In order to pursue faster growth and development of weaned piglets, increased dietary protein (CP) levels were favoured by the pig industry and the feed industry. The digestive organs of piglets were not fully developed at weaning, and the digestive absorption capacity of protein was limited. High-protein diets can cause allergic reactions in piglets, destroy intestinal structural integrity, reduce immunity, and cause intestinal flora imbalance. Undigested proteins were prone to produce toxic substances, such as ammonia and biogenic amines, after fermentation in the hindgut, which negatively affects the health of the intestine and eventually causes reduced growth performance and diarrhoea in piglets. This review revealed the mechanism of diarrhoea caused by high-protein diets in weaned piglets and provided ideas for preventing diarrhoea in weaned piglets.


Subject(s)
Animal Feed , Diet, High-Protein , Swine , Animals , Weaning , Animal Feed/analysis , Diet , Diarrhea/veterinary , Diet, High-Protein/veterinary , Dietary Supplements
7.
Braz. j. biol ; 82: 1-5, 2022. tab
Article in English | LILACS, VETINDEX | ID: biblio-1468548

ABSTRACT

For the proper growth of fish, it is necessary to feed the fish with a proper and balanced diet. A study was conducted to find out the effect of different protein-based diets on fingerlings of Tor putitora (mahseer). A feed with dietary protein levels of 35%, 40%, 45%, and 50% were prepared. The effect of different protein-based feed on weight gain, standard growth rate (SGR), food conversion ratio (FCR), percent weight gain, food conversion efficiency (FCE), and protein efficiency ratio (PER) was studied. An increase was observed in the growth rate with an increase in protein concentration up to 45%. The fingerlings fed a 45% protein diet shown the highest growth, followed by 50%, 40%, and 35%. The SGR value was greatest for 45% protein diet (8.56) followed by 50% and 40%, while the least values were observed for 35% protein feed (1.57). The same trend was observed for FCE. The highest PER values was observed in fishes fed 45% protein-based feed (0.65) followed by 50% (0.56), 40% (0.38) and35% (0.17). The food conversion ratio was lowest for 45% protein diet (3.41), while the greatest for 35% protein feed (16.85). It was concluded that a 45% protein-based diet was the best feed formulation for higher production of Tor putitora. However, research on the same percentage of protein diet is recommended for yearlings.


Para o bom crescimento dos peixes, é necessário alimentá-los com uma alimentação adequada e balanceada. Um estudo foi realizado para descobrir o efeito de diferentes dietas à base de proteínas em alevinos de Tor putitora (mahseer). Foi preparado um alimento com níveis de proteína dietética de 35%, 40%, 45% e 50%. O efeito de diferentes alimentos à base de proteína no ganho de peso, taxa de crescimento padrão (SGR), taxa de conversão alimentar (FCR), ganho de peso percentual, eficiência de conversão alimentar (FCE) e taxa de eficiência proteica (PER) foi estudado. Foi observado um aumento na taxa de crescimento com um aumento na concentração de proteína de até 45%. Os alevinos alimentados com dieta de 45% de proteína apresentaram o maior crescimento, seguidos de 50%, 40% e 35%. O valor de SGR foi maior para dieta com 45% de proteína (8,56), seguido de 50% e 40%, enquanto os menores valores foram observados para ração com 35% de proteína (1,57). A mesma tendência foi observada para FCE. Os maiores valores de PER foram observados em peixes alimentados com 45% de ração à base de proteína (0,65), seguido por 50% (0,56), 40% (0,38) e 35% (0,17). A taxa de conversão alimentar foi menor para a dieta com 45% de proteína (3,41), enquanto a maior para a dieta com 35% de proteína (16,85). Concluiu-se que a dieta à base de proteína de 45% foi a melhor formulação alimentar para maior produção de Tor putitora. No entanto, a pesquisa sobre a mesma porcentagem de dieta proteica é recomendada para animais de um ano.


Subject(s)
Animals , Cyprinidae/growth & development , Cyprinidae/metabolism , Diet, High-Protein/adverse effects , Diet, High-Protein/veterinary
8.
J Dairy Sci ; 104(5): 5601-5616, 2021 May.
Article in English | MEDLINE | ID: mdl-33663838

ABSTRACT

Increasing the supply of metabolizable protein (MP) and improving its AA profile may attenuate body protein mobilization in fresh cows and lead to increased milk production. Increasing the concentration of rumen-undegradable protein (RUP) to increase MP supply and replacing RUP sources from forages instead of nonforage fiber sources may further decrease tissue mobilization if it improves dry matter intake (DMI). Our objective was to determine whether increasing MP concentrations and improving the AA profile at the expense of either nonforage or forage fiber (fNDF) would affect MP balance and empty body (EB) composition (measured using the urea dilution method) in early postpartum dairy cows of different parities. In a randomized block design, 40 primigravid [77 ± 1.5 kg of EB crude protein (CP) at 8 ± 0.6 d before calving] and 40 multigravid (92 ± 1.6 kg of EB CP at 5 ± 0.6 d before calving) Holsteins were blocked by calving date and fed a common prepartum diet (11.5% CP). After calving to 25 d in milk (DIM), cows were fed 1 of 4 diets: (1) a diet deficient in MP meeting 87% of MP requirements (DMP, 17% CP, 24% fNDF), (2) 104% of MP requirements using primarily soy protein to make MP adequate (AMP, 20% CP, 24% fNDF), (3) 110% of MP requirements using a blend of proteins and rumen-protected (RP) AA to make MP adequate (Blend, 20% CP, 24% fNDF), or (4) a diet similar to Blend but substituting added RUP for fNDF rather than nonforage NDF (Blend-fNDF, 20% CP, 19% fNDF). Blend was formulated to have a RUP supply with a similar AA profile to that of casein. Cows were fed a common diet (16.3% CP) from 26 to 50 DIM. Calculated MP balance (supply - requirements) was less than zero for DMP and Blend-fNDF from 1 to 4 wk of lactation (WOL), whereas that for AMP was positive from 1 to 4 WOL and that for Blend was close to zero from 3 to 4 WOL. Daily MP balance was greater from 5 to 7 WOL for DMP compared with AMP and Blend (100 vs. 22 g/d). From -7 to 7 d relative to calving, losses of EB CP were greater for DMP than for AMP and Blend (-121 vs. average of 11 g/d). From 7 to 25 DIM, cows fed AMP (-139 g/d) and Blend-fNDF (-147 g/d) lost EB CP but cows fed Blend (-8 g/d) maintained EB CP. Increased DMI for Blend versus AMP led to reduced losses of EB lipid in primiparous cows from 7 to 25 d relative to calving (-1.0 vs. -1.3 kg/d of EB lipid), whereas lipid mobilization was similar in multiparous cows (average -1.1 kg of EB lipid/d). By 50 DIM, EB lipid and CP were similar across treatments and parities (average 60.2 kg of EB lipid and 81.6 kg of EB CP). Overall, feeding fresh cows a high MP diet with a balanced AA profile improved DMI and attenuated EB CP mobilization, which could partly explain positive carryover effects on milk production for multiparous cows and reduced lipid mobilization for primiparous cows.


Subject(s)
Amino Acids , Diet, High-Protein , Amino Acids/metabolism , Animals , Body Composition , Cattle , Diet/veterinary , Diet, High-Protein/veterinary , Dietary Proteins/metabolism , Female , Lactation , Milk/metabolism , Rumen/metabolism
9.
J Dairy Sci ; 104(5): 5583-5600, 2021 May.
Article in English | MEDLINE | ID: mdl-33663852

ABSTRACT

Because of low feed intake during the first weeks of lactation, dietary concentration of metabolizable protein (MP) must be elevated. We evaluated effects of providing additional rumen-undegradable protein (RUP) from a single source or a blend of protein and AA sources during the first 3 wk of lactation. We also evaluated whether replacing forage fiber (fNDF) or nonforage fiber with the blend affected responses. In a randomized block design, at approximately 2 wk prepartum, 40 primigravid (664 ± 44 kg of body weight) and 40 multigravid (797 ± 81 kg of body weight) Holsteins were blocked by calving date and fed a common diet (11.5% crude protein, CP). After calving to 25 d in milk (DIM), cows were fed 1 of 4 diets formulated to be (1) 20% deficient in metabolizable protein (MP) based on predicted milk production (17% CP, 24% fNDF), (2) adequate in MP using primarily RUP from soy to increase MP concentration (AMP; 20% CP, 24% fNDF), (3) adequate in MP using a blend of RUP and rumen-protected AA sources to increase MP concentration (Blend; 20% CP, 24% fNDF), or (4) similar to Blend but substituting fNDF with added RUP rather than nonforage neutral detergent fiber (Blend-fNDF; 20% CP, 19% fNDF). The blend was formulated to have a RUP supply with an AA profile similar to that of casein. A common diet (17% CP) was fed from 26 to 92 DIM, and milk production and composition were measured from 26 to 92 DIM, but individual dry matter intake (DMI) was measured only until 50 DIM. During the treatment period for both parities, AMP and Blend increased energy-corrected milk (ECM) yields compared with the diet deficient in MP based on predicted milk production (40.7 vs. 37.8 kg/d) and reduced concentrations of plasma 3-methyl-His (4.1 vs. 5.3 µmol/L) and growth hormone (9.0 vs. 11.9 ng/mL). Blend had greater DMI than AMP (17.4 vs. 16.1 kg/d), but ECM yields were similar. Blend had greater plasma Met (42.0 vs. 26.4 µmol/L) and altered metabolites associated with antioxidant production and methyl donation compared with AMP. Conversely, the concentration of total essential AA in plasma was less in Blend versus AMP (837 vs. 935 µmol/L). In multiparous cows, Blend-fNDF decreased DMI and ECM yield compared with Blend (19.2 vs. 20.1 kg/d of DMI, 45.3 vs. 51.1 kg/d of ECM), whereas primiparous cows showed the opposite response (15.3 vs. 14.6 kg/d of DMI, 32.9 vs. 31.4 kg/d of ECM). Greater DMI for multiparous cows fed Blend carried over from 26 to 50 DIM and was greater compared with AMP (23.1 vs. 21.2 kg /d) and Blend-fNDF (21.3 kg/d). Blend also increased ECM yield compared with AMP (49.2 vs. 43.5 kg/d) and Blend-fNDF (45.4 kg/d) from 26 to 92 DIM. Few carryover effects of fresh cow treatments on production were found in primiparous cows. Overall, feeding blends of RUP and AA may improve the balance of AA for fresh cows fed high MP diets and improve concurrent and longer-term milk production in multiparous cows. However, with high MP diets, multiparous fresh cows require greater concentrations of fNDF than primiparous cows.


Subject(s)
Amino Acids , Diet, High-Protein , Animals , Cattle , Diet/veterinary , Diet, High-Protein/veterinary , Dietary Proteins , Female , Lactation , Milk , Rumen
10.
J Dairy Sci ; 104(4): 4134-4145, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33612237

ABSTRACT

Feeding corn dried distillers grains with solubles (DDGS) in low crude protein (CP) diets could limit N waste in lactating cows. However, it also could possibly reduce metabolizable AA supply, especially Lys, and compromise milk production. Therefore, the objective of this study was to evaluate the effects of feeding supplemental blends of rumen undegradable protein (RUP) and rumen-protected (RP) AA in a low compared with high CP diet containing corn DDGS on milk production and selected measures of N utilization. Six multiparous Holstein cows (619.3 ± 49.8 kg of body weight; 26.8 ± 6.2 d in milk) were subjected to a split-plot, 3 × 3 Latin square design (21-d periods) with dietary CP content [low (14.6%; LP) or high (16.6%; HP)] as the whole-plot factor, and blend of RUP and RP-AA [control (CON), no supplement; blend A (0.11 kg/cow per d); or blend B (0.45 kg/cow per d)] as the sub-plot factor. All diets contained 10% corn DDGS; blends of RUP and RP-AA were top-dressed during morning feeding. There was no dietary CP content × supplemental blend interaction for all measured variables. Nutrient (dry matter, organic matter, neutral detergent fiber, acid detergent fiber, and CP), milk and milk component yields, and feed and apparent N efficiency did not differ for cows fed the low- compared with the high-protein diet. However, apparent total-tract CP digestibility, blood and milk urea-N concentrations, and urinary excretion (g/d) of N and urea-N were lower for cows fed the low-protein compared with the high-protein diet. There was no supplemental blend effect on nutrient intake and apparent total-tract digestibility, and milk and milk component yields. Except for a tendency for total urinary purine derivative excretion and microbial N flow to be lower for cows fed blend B compared with CON but not blend A, there was no supplemental blend effect on measures of N utilization. Both dietary CP content and supplemental blend of RUP and RP-AA had a marginal effect on the plasma free AA profile. In summary, reducing dietary CP content in diets containing corn DDGS had no effect on lactation performance, possibly accounting for the lack of a positive response following the provision of supplemental blends of RUP and RP-AA. However, reducing dietary CP content resulted in a decrease in blood and milk urea-N concentrations, and urinary excretion of N and urea-N, suggestive of an improvement in the efficiency of N use.


Subject(s)
Diet, High-Protein , Rumen , Amino Acids , Animal Feed/analysis , Animals , Cattle , Diet/veterinary , Diet, High-Protein/veterinary , Dietary Proteins , Female , Lactation , Milk/chemistry , Nitrogen/analysis , Rumen/chemistry , Zea mays
11.
PLoS One ; 16(2): e0247718, 2021.
Article in English | MEDLINE | ID: mdl-33630953

ABSTRACT

The objective of this study was to investigate the effect of diet crude protein (CP) content and metabolisable energy (ME) intake on skeletal growth and associated parameters of growing steers prior to and during compensatory growth in weight and catch-up growth in skeletal elongation. The experiment was a factorial design with two cattle genotypes [Brahman crossbred (BX, 178 ± 6 kg) and Holstein-Friesian (HF, 230 ± 34 kg)] and three nutritional treatments; high CP content and high ME intake (HCP-HME), high CP content and low ME intake (HCP-LME) and low CP content and low ME intake (LCP-LME) with the ME intake of HCP-LME matched to that of LCP-LME. Nutritional treatments were imposed over a 103 d period (Phase 1), and after this, all steers were offered ad libitum access to the HCP-HME nutritional treatment for 100 d (Phase 2). Steers fed the high CP content treatment with a low ME intake, showed higher hip height gain (P = 0.04), larger terminal hypertrophic chondrocytes (P = 0.02) and a higher concentration of total triiodothyronine in plasma (P = 0.01) than steers with the same ME intake of the low CP content treatment. In addition, the low CP treatment resulted in significant decreases in bone volume (P = 0.03), bone surface area (P = 0.03) and the concentration of bone-specific alkaline phosphatase in plasma (P < 0.001) compared to steers fed the HCP-HME treatment. A significant interaction between genotype and nutritional treatment existed for the concentration of thyroxine (T4) in plasma where HF steers fed LCP-LME had a lower T4 concentration in plasma (P = 0.05) than BX steers. All steers with a restricted ME intake during Phase 1 demonstrated compensatory growth during Phase 2. However, HF steers fed the LCP treatment during Phase 1 showed a tendency (P = 0.07) for a greater LWG during Phase 2 without any increase in dry matter intake. Results observed at the growth plate and hip height growth suggest that catch-up growth in cattle may also be explained by the growth plate senescence hypothesis. Contrary to our initial hypothesis, the results demonstrate that greater CP intake during ME restriction does not increase compensatory gain in cattle during re-alimentation.


Subject(s)
Animal Feed , Animal Nutritional Physiological Phenomena , Bone and Bones/metabolism , Diet, High-Protein/veterinary , Dietary Proteins/administration & dosage , Energy Intake , Animals , Bone Development , Cattle , Energy Metabolism , Genotype , Male
12.
Metabolomics ; 16(10): 114, 2020 10 12.
Article in English | MEDLINE | ID: mdl-33047270

ABSTRACT

INTRODUCTION: Ketoacidosis of metabolic disease showed in beef cattle although body weight was increased by high-grain diets (HGDs). However, few studies have examined for health status related with metabolic disease of ketoacidosis following high-protein diet (HPD). OBJECTIVES: Metabolomic analysis was performed for the monitoring of health status associated with metabolic disease of ketoacidosis in the plasma of Hanwoo heifers following a HPD. METHODS: Hanwoo heifers of 24 months with 459 ± 42 kg weight were used under a 2 × 2 crossover design. The plasma was collected from control (n = 5) and HPD group (n = 5) on day 21 following diet adaptation for 20 days. Metabolic profiling analysis of organic acids (OAs), amino acids (AAs) and fatty acids (FAs) by gas chromatography-tandem mass spectrometry combined with star pattern analysis was performed in plasma. Levels of OAs, AAs and FAs were evaluated by Mann-Whitney test, PCA and PLS-DA. RESULTS: In HPD group, ketoacidosis as metabolic disease was monitored by elevated acetoacetic acid and 3-hydroxybutyric acid. In addition, the elevation of ketogenic AAs, reduction of medium chain FAs and OAs with energy metabolism in TCA cycle were monitored in HPD group. Star graphic pattern was characteristic and readily distinguished between control and HPD groups. In PLS-DA, two groups were separated with VIP score of top-ranked 10 FAs as important metabolites for discrimination. CONCLUSION: Elevation of ketone body including ketogenic AAs and reduced energy metabolism of FAs and OAs may useful for evaluation of health states associated with ketoacidosis from metabolic event by HPD in beef cattle.


Subject(s)
Amino Acids/blood , Cattle/blood , Ketosis/blood , Animals , Diet, High-Protein/adverse effects , Diet, High-Protein/veterinary , Fatty Acids/blood , Female , Gas Chromatography-Mass Spectrometry/methods , Ketosis/diagnosis , Metabolomics/methods , Republic of Korea
13.
J Vet Intern Med ; 34(2): 852-856, 2020 Mar.
Article in English | MEDLINE | ID: mdl-32086838

ABSTRACT

BACKGROUND: Dietary protein and amino acid intake and composition can modulate immune function. OBJECTIVES: To evaluate the effects of high-protein intake and arginine and ornithine supplementation on feline immune cells. ANIMALS: Ten healthy cats. METHODS: Experimental study. Cats received a high-protein basal diet as a single daily meal. A crossover design was applied with treatments being basal diet (w/o); basal diet with arginine supplementation (+50, 75, 100% compared to the arginine provision by the basal diet; Arg 1-3); and basal diet with ornithine supplementation (+100, 150, 200% compared to the arginine provision by the basal diet; Orn 1-3). Blood samples were collected at the end of each 11-day treatment period. RESULTS: Mitogen-stimulated proliferative activity of blood leukocytes revealed a quadratic effect for the dietary supplementation of arginine (P = .02) and ornithine (P = .03) (means for ConA-stimulation: w/o = 6.96; Arg 1 = 9.31; Arg 2 = 11.4; Arg 3 = 8.04; Orn 1 = 15.4; Orn 2 = 9.43; Orn 3 = 9.28; pooled SEM: 0.96). The number (% gated) of phagocytic granulocytes linearly decreased with increasing dietary concentrations of arginine (P = .05) and ornithine (P = .03) (means: w/o = 95.5; Arg 1 = 93.0; Arg 2 = 92.5; Arg 3 = 92.6; Orn 1 = 92.6; Orn 2 = 92.6; Orn 3 = 91.5; pooled SEM = 0.44). CONCLUSIONS AND CLINICAL IMPORTANCE: This study could demonstrate immunomodulating properties of dietary arginine and ornithine in cats.


Subject(s)
Arginine/administration & dosage , Cats/immunology , Diet, High-Protein/veterinary , Immunity, Cellular , Leukocytes/immunology , Ornithine/administration & dosage , Animal Nutritional Physiological Phenomena , Animals , Dietary Supplements , Female , Male
14.
J Anim Sci ; 97(12): 4865-4874, 2019 Dec 17.
Article in English | MEDLINE | ID: mdl-31679024

ABSTRACT

Vitamin B6 (VB6) is an important coenzyme factor which participates in many metabolic reactions, especially amino acid metabolism. There are few reports on how VB6 mediates weaned piglet intestinal health. This study purposed to investigate dietary VB6 effects on growth, diarrhea rates, and intestinal morphology and function in weaned piglets fed a high-crude protein (22% CP) diet. Eighteen 21-d-old weaned [(Yorkshire × Landrace) × Duroc] piglets with body weights of 7.03 ± 0.15 (means ± SEM) kg were randomly assigned into 3 VB6-containing dietary treatments. Vitamin B6 content was: 0, 4, and 7 mg/kg, respectively. The feeding period lasted 14 d. The results showed that no significant difference existed for the growth performance. The 7 mg/kg VB6 group had a tendency to decrease diarrhea rate (P = 0.065). Blood biochemical parameters analysis demonstrated that total protein, cholesterol, and high-density lipoprotein significantly increased in the 7 mg/kg VB6 group (P < 0.05). In the jejunum, no significant differences were detected for villus height, villus width, crypt depth, villus height and crypt depth ratios, and positive Ki67 counts and the mRNA expression of inflammatory cytokines. Vitamin B6 significantly increased the mRNA expression of SLC6A19 and SLC6A20 (P < 0.05) and decreased the mRNA expression of SLC36A1 (P < 0.05). In the ileum, VB6 significantly increased villus height and villus width (P < 0.05) while decreased positive Ki67 cell counts for 7 mg/kg VB6 group (P < 0.05). Vitamin B6 had significantly increased the mRNA expression of interleukin-1ß, tumor necrosis factor-α,cyclo-oxygen-ase-2, and transforming growth factor-ß (P < 0.05). Vitamin B6 also had significantly increased mRNA expression of SLC6A19, SLC7A6, SLC7A7, and SLC36A1 (P < 0.05). These findings suggest that dietary supplementation with VB6 may affect the intestinal morphology and absorption and metabolism of protein in weaned piglets fed a high-protein diet by altering the expression of intestinal inflammatory cytokines and amino acid transporters.


Subject(s)
Diarrhea/veterinary , Diet, High-Protein/veterinary , Dietary Supplements/analysis , Swine/physiology , Vitamin B 6/administration & dosage , Animals , Cytokines/analysis , Diet/veterinary , Immunohistochemistry/veterinary , Intestinal Mucosa/cytology , Intestinal Mucosa/drug effects , Intestines/anatomy & histology , Intestines/cytology , Intestines/drug effects , Weaning
15.
Arch Anim Nutr ; 73(4): 306-323, 2019 Aug.
Article in English | MEDLINE | ID: mdl-31164000

ABSTRACT

Diets excessive in crude protein (CP) are unfavourable in terms of metabolic and environmental load. Dietary phenols, often binding to dietary proteins, may alleviate these problems. In an experiment with 60 lambs (3.2 ± 1.6 months of age; 29.7 ± 5.1 kg body weight), kept in pairs, five diets were tested. A diet with 157 g CP/kg dry matter (DM) served as negative control. Four diets with on average 229 (225-233) g CP/kg DM remained either non-supplemented or were supplemented with 13 g/kg DM of Acacia mearnsii extract, grapeseed extract, or a combination of both (26 g extract/kg DM). The analysed concentrations of total extractable phenols were 7.1, 8.1, 14.3, 16.6 and 25.4 g/kg DM for low (CP‒) and high CP (CP+), and high CP with acacia (CP+A), grapeseed (CP+G) and acacia plus grapeseed (CP+AG), respectively. Diets were fed for 10 weeks, and for 6 d faeces and urine were collected and subsequently stored as complete manure for 8 weeks. In blood plasma, phenol concentrations and activities of enzymes indicating liver and kidney stress were analysed. The CP+ diet increased apparent digestibility of N and its removal with the urine, with the expected increase in gaseous N emissions from the manure (13.5 vs 6.5 g/lamb per day during 8 weeks) compared to CP‒. However, no clear signs of metabolic stress were detected. Supplementing the extracts did not impair intake, growth performance and digestibility. Only the supplementation with both extracts decreased urinary N proportion of manure N, and the concomitant weak decline in gaseous emission from the manure was not significant. At least part of the phenols of both extracts seem to be bioavailable as their supplementation elevated blood plasma phenol concentrations by 15% to 40% compared to CP+. A combination of both extracts did not result in a further increase. Further studies have to identify the minimally effective dosage for reducing N emissions, which, at the same time, does not cause adverse side effects in performance.


Subject(s)
Acacia/chemistry , Diet, High-Protein/veterinary , Manure/analysis , Nitrogen/metabolism , Plant Extracts/pharmacology , Sheep, Domestic/metabolism , Air Pollutants/analysis , Animal Feed/analysis , Animals , Diet/veterinary , Dietary Supplements/analysis , Female , Grape Seed Extract/pharmacology , Male , Random Allocation
16.
Arch Anim Nutr ; 73(5): 399-413, 2019 Oct.
Article in English | MEDLINE | ID: mdl-31137970

ABSTRACT

This in vitro study aimed to evaluate the effects of Yucca schidigera powder (YSP) and inulin (IN) on protein fermentation metabolites (short-chain fatty acids [SCFA] and branched-chain fatty acids, phenolic and indolic compounds, biogenic amines, ammonia and pH) by using faecal inocula from dogs fed either a low (L) or a high (H) protein diet (crude protein 201 or 377 g/kg as fed). Four treatments for each diet were evaluated in an in vitro batch culture system over 24 h: (1) control with no addition of substrates; (2) 4 g YSP/l; (3) 5 g IN/l; (4) 4 g YSP/l in combination with 5 g IN/l of faecal culture of dogs fed Diet L or H. Several changes in fermentation metabolites were analysed. Samples incubated with the faecal inocula of dogs fed Diet L produced higher concentrations of total SCFA, propionate (p = 0.001), acetate (p ≤ 0.001), d-lactate (p = 0.041) and indole (p = 0.003), whereas pH (p = 0.004) was decreased. Supplementation of IN increased the content of putrescine, d- and l-lactate, total SCFA, acetate, propionate, n-butyrate (p ≤ 0.001) and n-valerate (p = 0.003), while i-valerate, indole and pH (p ≤ 0.001) were reduced. Ammonia was lower (p = 0.013) in samples with faecal inocula from dogs fed Diet H and further reduced by the addition of IN (p ≤ 0.001). Samples with faecal inocula from dogs fed Diet L had a fewer quotient of ammonia and total SCFA (p = 0.040). Supplementation of YSP (p = 0.021), IN (p ≤ 0.001) and YSP in combination with IN (p = 0.047) led to a higher reduction of the quotient of ammonia and total SCFA. In conclusion, dietary protein concentration and the supplementation of IN resulted in a stimulation of fermentation while YSP appeared to have only minor effects.


Subject(s)
Diet, High-Protein/veterinary , Diet, Protein-Restricted/veterinary , Dietary Proteins/metabolism , Feces/microbiology , Inulin/metabolism , Yucca/chemistry , Animal Feed/analysis , Animal Nutritional Physiological Phenomena/drug effects , Animals , Diet/veterinary , Dietary Supplements/analysis , Dogs , Female , Fermentation/drug effects , In Vitro Techniques , Inulin/administration & dosage , Male
17.
PLoS One ; 14(5): e0216072, 2019.
Article in English | MEDLINE | ID: mdl-31042730

ABSTRACT

Commercial diets high in animal protein and fat are increasingly being developed for pets, however little is understood about the impacts of feeding such diets to domestic cats. The carbohydrate content of these diets is typically low, and dietary fibre is often not included. Dietary fibre is believed to be important in the feline gastrointestinal tract, promoting stool formation and providing a substrate for the hindgut microbiome. Therefore, we aimed to determine the effects of adding plant-based dietary fibre to a high animal protein and fat diet. Twelve domestic short hair cats were fed three complete and balanced diets in a cross-over design for blocks of 21 days: raw meat (Raw), raw meat plus fibre (2%, 'as is' inclusion of inulin and cellulose; Raw+Fibre) and a commercially available Kibble diet. A commercially available canned diet was fed for 21 days as a washout phase. Apparent macronutrient digestibility, faecal output, score, pH, organic acid concentrations and bacteriome profiles were determined. Diet significantly affected all faecal parameters measured. The addition of dietary fibre to the raw meat diet was found to reduce apparent macronutrient digestibility, increase faecal output, pH and score. Thirty one bacterial taxa were significantly affected by diet. Prevotella was found to dominate in the Kibble diet, Clostridium and Fusobacterium in the Raw diet, and Prevotella and a group of unclassified Peptostreptococcaceae in the Raw+Fibre diet. Our results show that diets of different macronutrient proportions can strongly influence the faecal microbiome composition and metabolism, as shown by altered organic acid concentrations and faecal pH, in the domestic cat. The addition of 2% of each fibre to the Raw diet shifted faecal parameters closer to those produced by feeding a Kibble diet. These results provide a basis for further research assessing raw red meat diets to domestic cats.


Subject(s)
Animal Feed/analysis , Diet, High-Fat/veterinary , Diet, High-Protein/veterinary , Animal Nutritional Physiological Phenomena , Animals , Cats/metabolism , Diet/veterinary , Dietary Fiber/analysis , Feces/chemistry , Female , Male , Meat/analysis , Microbiota/physiology , Red Meat
18.
Sci Rep ; 9(1): 4822, 2019 03 18.
Article in English | MEDLINE | ID: mdl-30886210

ABSTRACT

Obesity and inactivity are major risk factors of feline diabetes mellitus (FDM) and human type II diabetes mellitus (T2DM). In recent years, changes in the gut microbiota have been suggested as a contributing factor to T2DM. Whether the gut microbiota (GM) composition plays a role in FDM remains unknown. The aim of the current study was firstly a cross-sectional comparison of the GM of diabetic cats, to that of lean, and of obese/overweight non-diabetic cats of a similar age. Specifically, fecal samples from 82 privately-owned cats from Denmark and Switzerland were sequenced using 16S rRNA gene amplicon metabarcoding. Secondly dietary intervention data was generated, by obtaining additional samples from a subset of cats after placing them on a high-protein diet for four weeks. The GM diversity of diabetic cats was lower than that of lean cats in the cross-sectional study, and lower compared to lean and to overweight/obese cats after diet intervention. Diabetic cats also exhibited fewer Anaerotruncus, Dialister, and unknown Ruminococcaceae than lean cats. Serum fructosamine levels correlated negatively with Prevotellaceae abundance and positively with Enterobacteriaceae abundance. In summary the intestinal microbiota of diabetic cats was characterized by decreased GM diversity and loss of butyrate producing bacterial genera.


Subject(s)
Bacteria/isolation & purification , Cat Diseases/microbiology , Diabetes Mellitus/veterinary , Dysbiosis/veterinary , Gastrointestinal Microbiome/physiology , Obesity/veterinary , Animals , Bacteria/classification , Bacteria/genetics , Bacteria/metabolism , Butyrates/metabolism , Cat Diseases/blood , Cat Diseases/diet therapy , Cat Diseases/metabolism , Cats , Cross-Sectional Studies , DNA Barcoding, Taxonomic , DNA, Bacterial/isolation & purification , Denmark , Diabetes Mellitus/diet therapy , Diabetes Mellitus/metabolism , Diabetes Mellitus/microbiology , Diet, High-Protein/veterinary , Dysbiosis/blood , Dysbiosis/metabolism , Dysbiosis/microbiology , Feces/microbiology , Female , Fructosamine/blood , Fructosamine/metabolism , Male , Obesity/diet therapy , Obesity/metabolism , Obesity/microbiology , Prospective Studies , RNA, Ribosomal, 16S/genetics , Switzerland
19.
J Anim Sci ; 97(5): 2087-2098, 2019 Apr 29.
Article in English | MEDLINE | ID: mdl-30901031

ABSTRACT

The objective of this study was to evaluate the effects of feeding high-protein corn distillers grains on fresh beef quality. Steers (n = 60) were fed one of five diets for 190 d (six pens with 10 cattle/pen/diet): a corn control (Control), 40% high-protein dry distillers grains plus solubles (HP-DDGS), 40% dry distillers grains plus solubles (DDGS), 40% wet distillers grains plus solubles (WDGS), or 40% bran plus solubles (Bran). Eighteen Choice carcasses (three cattle per pen) were randomly selected within each treatment; however, two carcasses were lost during carcass selection. Strip loins (Longissimus lumborum, IMPS # 180) were collected, divided into three equal portions, and aged for 2, 9, or 23 d. Steaks were fabricated following each aging period and placed under retail display (RD) conditions for 0, 4, or 7 d. Pen was the experimental unit. Hot carcass weight at harvest was 391 kg (SD 31.6 kg). Dietary treatment had no effect on tenderness (Warner-Bratzler shear force) within each aging period (P > 0.05). After 7 d of RD, following all aging periods, steaks from cattle fed HP-DDGS had the greatest visual discoloration except for DDGS and Bran after 23 d (P < 0.05). The steaks from cattle fed HP-DDGS had lower (P < 0.05) redness (a*) values than all other treatments following 7 d of RD. Lipid oxidation increased (P < 0.001) during RD at all aging periods, as measured via thiobarbituric acid reactive substance (TBARS). A day of RD-by-dietary treatment effect (P < 0.001) was observed for lipid oxidation. After 7 d of RD, steaks from cattle fed HP-DDGS had higher TBARS than all other treatments except WDGS (P < 0.01). A trend was found for sarcomere length (P = 0.07), with steaks from cattle fed Bran having the longest sarcomere length compared with all other treatments. An aging effect was found for free calcium content (P < 0.001) where steaks aged 9 and 23 d had significantly higher levels of free calcium than those aged 2 d. There was a tendency (P = 0.07) that steaks from cattle fed WDGS contained more free calcium than steaks from cattle fed DDGS or Bran. Although there were no statistical differences, there was a trend (P = 0.07) that steaks from cattle fed HP-DDGS had higher levels of PUFAs and C18:2. These results suggest that feeding high protein distillers grains has no detriment on tenderness, but may alter the lipid profile of the muscle, resulting in decreased color stability, increased lipid oxidation, and decreased shelf life.


Subject(s)
Animal Feed/analysis , Cattle/physiology , Diet, High-Protein/veterinary , Grain Proteins/metabolism , Red Meat/standards , Zea mays , Animals , Color , Diet/veterinary , Edible Grain , Ethanol , Fatty Acids, Unsaturated/analysis , Male
20.
Avian Dis ; 63(3): 461-467, 2019 09 01.
Article in English | MEDLINE | ID: mdl-31967429

ABSTRACT

Clostridium perfringens (CP) type A and newly created type G strains are the key etiological factors in the induction of necrotic enteritis (NE), an important enteric disease that is responsible for the annual loss of $6 billion in the worldwide poultry industry. Several CP toxin genes were found to be critical in NE pathogenesis in chickens, but limited information is available on the CP lethal toxin tpeL gene. In this study, 19 CP strains isolated from NE-affected chicken farms were characterized microbiologically and molecularly and evaluated for their pathogenicity in commercial broiler chickens. Toxin typing by PCR revealed that all strains tested were positive for the netB toxin gene, but only five strains were positive for the tpeL toxin gene (LLY-TpeL 13, -TpeL 15, -TpeL 17, -TpeL 18, and -TpeL 19, simplified as TpeL 13, TpeL 15, TpeL 17, TpeL 18, and TpeL 19). High levels of TpeL proteins were detected in the concentrated culture supernatant from strains TpeL 13, 15, 17, and 19 by western blotting. Quantitative PCR showed that strains TpeL 13, 15, 17, 18, and 19 harbored a high number of copies of tpeL genes, while TpeL 18 had the highest number of copies of the tpeL gene among all CP strains tested when normalized with copy numbers of 16S rRNA gene as a housekeeping gene marker. The in vivo NE challenge test using multiple oral CP inoculations combined with a high-protein diet showed that TpeL 17 was the most virulent in inducing typical NE lesions, followed by TpeL 19 as the next most virulent, when tested in commercial broiler chickens. Infection with TpeL 17 reduced the growth rate significantly, as shown by reduced relative body weight gain percentage at day 5 postinfection. Availability of the virulent netB+tpeL+ CP strains is essential for the development of a CP-alone NE challenge model that could provide significant tools for understanding CP pathogenesis and for development of alternative to antibiotics.


Caracterización de cepas virulentas de Clostridium perfringens netB+/tpeL+ de granjas de pollos de engorde con enteritis necrótica. Clostridium perfringens (CP) tipo A y los tipos nuevos G son los factores etiológicos clave en la inducción de enteritis necrótica (NE), una enfermedad entérica importante que es responsable de pérdidas anuales de $6 mil millones en la industria avícola mundial. Se ha determinado que varios genes de toxinas de C. perfringens son críticos en la patogénesis de la enteritis necrótica en pollos, pero se dispone de información limitada sobre el gene tpeL de la toxina letal de C. perfringens. En este estudio, se caracterizaron microbiológicamente y molecularmente 19 cepas de C. perfringens aisladas de granjas avícolas afectadas por enteritis necrótica y se evaluaron en su patogenicidad en pollos de engorde comerciales. La tipificación de toxinas por PCR reveló que todas las cepas analizadas fueron positivas para el gene de la toxina netB, pero solo cinco fueron positivas para el gene de la toxina tpeL (LLY-TpeL 13, -TpeL 15, -TpeL 17, -TpeL 18, y -TpeL 19, simplificado como TpeL 13, TpeL 15, TpeL 17, TpeL 18 y TpeL 19). Los altos niveles de proteínas TpeL se detectaron en el sobrenadante del cultivo concentrado de las cepas TpeL 13, 15, 17 y 19 mediante inmunoelectrotransferencia tipo Western. La PCR cuantitativa mostró que las cepas TpeL 13, 15, 17, 18 y 19 albergaban un alto número de copias de los genes tpeL, mientras que TpeL 18 mostró el mayor número de copias del gene tpeL entre todas las cepas de C. perfringens analizadas cuando se normalizó con los números de copias del gene 16S rRNA como un marcador genético constitutivo. La prueba in vivo de desafío de enteritis necrótica utilizando múltiples inoculaciones orales de C. perfringens combinadas con una dieta alta en proteínas mostró que TpeL 17 fue la más virulenta en la inducción de lesiones típicas de enteritis necrótica, seguida de TpeL 19 como la siguiente más virulenta cuando se inoculó en pollos de engorde comerciales. La infección con TpeL 17 redujo significativamente la tasa de crecimiento, como lo demuestra la reducción del porcentaje de aumento de peso corporal relativo en el día cinco posterior a la infección. La disponibilidad de las cepas netB+ tpeL+ de C. perfringens virulentas es esencial para el desarrollo de un modelo de desafío enteritis necrótica únicamente con C. perfringens que podría proporcionar herramientas significativas para comprender la patogénesis de C. perfringens y para el desarrollo de alternativas a los antibióticos.


Subject(s)
Chickens , Clostridium Infections/veterinary , Clostridium perfringens/genetics , Clostridium perfringens/pathogenicity , Enteritis/veterinary , Necrosis/veterinary , Poultry Diseases/microbiology , Animals , Clostridium Infections/microbiology , Diet, High-Protein/veterinary , Enteritis/microbiology , Necrosis/microbiology , RNA, Bacterial/analysis , RNA, Ribosomal, 16S/analysis , Virulence
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