Biochemical profile of heifers with spontaneous humeral fractures suggest that protein-energy malnutrition could be an important factor in the pathology of this disease.

CASE HISTORY: Serum and liver samples from 35, 2-year-old dairy heifers that had fractured one or both humeri post-calving between July and December 2019 were submitted to a diagnostic laboratory for analysis. Serum samples were analysed for albumin, ß-hydroxybutyrate (BHB), creatinine, Ca, Mg, phosphate, non-esterified fatty acids (NEFA), and serum Cu concentration. Liver samples were analysed for liver Cu concentration. Data were compared to published reference intervals. Data values for heifers that prior to fracture had grazed fodder beet were also compared to values for those that had grazed pasture. CLINICAL FINDINGS: Sixty-nine percent of heifers with humeral fracture had serum creatinine concentrations below the lower value of the reference range (55-130 µmol/L). In 3/32 (9%) heifers, serum NEFA concentrations were increased above the reference value indicating body fat mobilisation (≥1.2 mmol/L for peri-partum cows) and in 20/35 (57%) heifers BHB serum concentrations were above the reference value indicating subclinical ketosis (≥1.1 mmol/L for peri-partum cows). In 24/35 (69%) heifers, liver Cu concentration was low (≤ 44 µmol/kg) or marginal (45-94 µmol/kg). The concentration of Cu in serum was low (≤ 4.5 µmol/L) in 2/33 (6%) heifers and marginal (4.6-7.9 µmol/L) in 5/33 (15%) heifers. There was moderate positive correlation between the logged concentrations of Cu in paired liver and serum samples, r(31) = 0.43; (95% CI = 0.1-0.79; p = 0.014). One heifer had a serum phosphate concentration below the lower limit of the reference range (< 1.10 mmol/L). For all heifers, the concentrations of albumin, Ca, and Mg in serum were within the reference intervals (23-38 g/L, 2.00-2.60 mmol/L, and 0.49-1.15 mmol/L respectively). Over winter, 15/35 (43%) heifers grazed predominantly pasture, 14/35 (40%) grazed fodder beet and 6/35 (17%) had a mixed diet. CLINICAL RELEVANCE: In some of these heifers with humeral fractures, there was evidence for protein and/or energy malnutrition in the form of elevated NEFA and BHB concentrations and low creatinine concentrations in serum. Liver Cu concentrations were also reduced in most affected heifers. However, the absence of a control group means it is not possible to determine if these are risk factors for fracture or features common to all periparturient heifers. Clinical trials and molecular studies are needed to determine the true contribution of Cu and protein-energy metabolism to the pathogenesis of spontaneous humeral fractures in dairy heifers. ABBREVIATIONS: BHB: ß-hydroxybutyrate; NEFA: Non-esterified fatty acids.

Quantitative histologic evaluation reveals different degree of liver atrophy in cachectic and starved dogs.

Cases of neglect in dogs are among the forensic cases submitted most commonly for postmortem examination. Starvation is a form of primary protein-energy malnutrition in which the availability of food is severely restricted or absent; cachexia is a form of protein-energy malnutrition secondary to progressive metabolic derangement during chronic diseases. Despite both conditions leading to an emaciated appearance of the cadaver, discrimination between the two is crucial in forensic cases. We hypothesized that among emaciated dogs, the degree of liver atrophy in starved animals is higher than in cachectic ones, and that this can be investigated microscopically, regardless of the degree of cadaver decomposition. We studied 46 animals: 23 starved, 11 cachectic, and 12 control dogs. Portal tracts were identified by the presence of a bile duct and associated vascular structures recognizable by a thin rim of collagen still visible regardless of the degree of cadaver decomposition. The number of portal tracts per lpf (10×) was used as an indirect measure of atrophy. The number of portal tracts in starved dogs was significantly higher (p < 0.01) compared to both cachectic and control dogs, indicating a higher degree of liver atrophy in starvation. Measuring the density of portal tracts offers a reliable additional tool for discrimination between starvation and cachexia.

Análise morfométrica e quantitativa dos neurônios do plexo mientérico do ceco de ratos submetidos a intensa carência de proteína / Análisis morfométrica y cuantitativa de las neuronas del plexo mientérico del ceco de ratas sometidos a una intensa carencia de proteína / Quantitative and morphometric analysis of myenteric neuron from the cecum of rats submitted to intensive protein deficiency

Avaliaram-se os efeitos crônicos de uma dieta severamente hipoprotéica, sobre aspectos quantitativos e morfométricos de neurônios mientéricos, NADPH-diaforase positivos do ceco de ratos jovens. Oito ratos foram divididos em dois grupos, sendo que para um dos grupos foi oferecida ração com teor protéico de 26% (controle) e, para o outro, ração com o teor protéico de 4%, mantendo-se o balanço vitamínico e mineral durante 12 semanas. Após eutanásia, o ceco de cada animal foi coletado e dissecado para confecção de preparados de membrana, os quais foram submetidos à técnica da NADPH-diaforase. Para contagem e mensuração destes neurônios, o ceco foi dividido em quatro regiões: apical mesentérica; apical antimesentérica; basal antimesentérica; próxima à ampola cecal. Não se observou alteração significativa para o peso dos animais, área do ceco e densidade populacional dos neurônios mientéricos no final do experimento. Por outro lado, em algumas regiões do ceco, os neurônios tiveram redução da área nuclear, com conseqüente redução da área do pericário

The chronicle effects of a severe hypoproteic diet on quantitative and morphometric aspects of NADPHdiaphorase-positive myenteric neurons from the cecum of young rats were assessed. Eight rats were divided into two groups. One group (control) was fed with 26%-protein chow and the other with 4%-protein chow during 12 weeks. The vitamin and mineral balance was kept. After euthanasia, the cecum of each animal was collected and dissected for the whole-mount preparation, which were submitted to NAPDH-diaphorase. The cecum was divided into four regions for counting and measurement of neurons: apical mesenteric, antimesenteric apical, basal antimesenteric, and next to the cecal ampulla. Significant alterations were not noted with respect to the weight of the animals, cecum area, and population density of myenteric neurons at the end of the experiment. On the other hand, in some regions of the cecum, the neurons presented a reduction of the nuclear area with resulting reduction of the soma area

Se evaluaron los efectos crónicos de una dieta severamente hipoprotéica, sobre aspectos cuantitativos y morfométricos de neuronas mientéricos, NADPH-diaforase positivos del ceco de ratas jóvenes. Ocho ratas fueron divididas en dos grupos, siendo que para uno de los grupos fue ofrecida ración con tenor proteico de 26% (control) y, para el otro, ración con el tenor proteico de 4%, manteniéndose el balance vitamínico y mineral durante 12 semanas. Tras eutanasia, El ceco de cada animal fue recolectado y disecado para confección de preparados de membrana, los cuales fueron sometidos a la técnica de la NADPH-diaforase. Para recuento y mensuración de estas neuronas, el ceco fue dividido en cuatro regiones: apical mesentérica, apical antimesentérica, basal antimesentérica y próxima a la ampolla cecal. No se observó alteración significativa en el peso de los animales, área del ceco y densidad poblacional de las neuronas mientéricas al final Del experimento. Por otro lado, en algunas regiones del ceco las neuronas tuvieron reducción del área nuclear, con consecuente reducción del área del pericario

Morphometric and ultrastructural analysis of different pituitary cell populations in undernourished monkeys.

Undernutrition elicited by a low-protein diet determines a marked reduction of hypophyseal activity and affects the function of the respective target organs. The objective of the present investigation was to study the ultrastructural and quantitative immunohistochemical changes of the different pituitary cell populations in undernourished monkeys that had been previously shown to have significant changes in craniofacial growth. Twenty Saimiri sciureus boliviensis monkeys of both sexes were used. The animals were born in captivity and were separated into two groups at one year of age, i.e., control and undernourished animals. The monkeys were fed ad libitum a 20% (control group) and a 10% (experimental group) protein diet for two years. Pituitaries were processed for light and electron microscopy. The former was immunolabeled with anti-GH, -PRL, -LH, -FSH, -ACTH, and -TSH sera. Volume density and cell density were measured using an image analyzer. Quantitative immunohistochemistry revealed a decrease in these parameters with regard to somatotrophs, lactotrophs, gonadotrophs and thyrotrophs from undernourished animals compared to control ones. In these populations, the ultrastructural study showed changes suggesting compensatory hyperfunction. On the contrary, no significant changes were found in the morphometric parameters or the ultrastructure of the corticotroph population. We conclude that in undernourished monkeys the somatotroph, lactotroph, gonadotroph, and thyrotroph cell populations showed quantitative immunohistochemical changes that can be correlated with ultrastructural findings.

Morphometric and ultrastructural analysis of different pituitary cell populations in undernourished monkeys

Undernutrition elicited by a low-protein diet determines a marked reduction of hypophyseal activity and affects the function of the respective target organs. The objective of the present investigation was to study the ultrastructural and quantitative immunohistochemical changes of the different pituitary cell populations in undernourished monkeys that had been previously shown to have significant changes in craniofacial growth. Twenty Saimiri sciureus boliviensis monkeys of both sexes were used. The animals were born in captivity and were separated into two groups at one year of age, i.e., control and undernourished animals. The monkeys were fed ad libitum a 20 percent (control group) and a 10 percent (experimental group) protein diet for two years. Pituitaries were processed for light and electron microscopy. The former was immunolabeled with anti-GH, -PRL, -LH, -FSH, -ACTH, and -TSH sera. Volume density and cell density were measured using an image analyzer. Quantitative immunohistochemistry revealed a decrease in these parameters with regard to somatotrophs, lactotrophs, gonadotrophs and thyrotrophs from undernourished animals compared to control ones. In these populations, the ultrastructural study showed changes suggesting compensatory hyperfunction. On the contrary, no significant changes were found in the morphometric parameters or the ultrastructure of the corticotroph population. We conclude that in undernourished monkeys the somatotroph, lactotroph, gonadotroph, and thyrotroph cell populations showed quantitative immunohistochemical changes that can be correlated with ultrastructural findings

Cystic fibrous osteodystrophy of the jaw in two pet kinkajous (Potos flavus).

Two kinkajous (Potos flavus), which were separately owned as pets and fed an inappropriate diet consisting almost exclusively of succulent fruits developed cystic fibrous osteodystrophy of the jaw. In both cases there was a prominent enlargement of the chin and lower jaw, decalcification of facial bones, softening of the gingiva, and severe malocclusion, but no other relevant changes. Clinical, radiographic and histological findings were consistent with cystic fibrous osteodystrophy. The findings suggest that kinkajous with this condition are particularly prone to developing a bulbous enlargement of the chin as the main clinical sign, and that this change is histologically similar to the cystic form of the disease observed in man and monkeys.

Effect of dietary restriction on cell-mediated immune responses in cattle infected with Mycobacterium bovis.

Nine M. bovis-infected cattle on a diet deficient in both protein and energy for 133 days lost approximately 17% of their original body weight. However, dietary restriction did not result in any significant reduction in skin sensitivity to PPD, in vitro production of IFN-gamma or lymphocyte blastogenesis. The number of circulating BoCD4+ cells and B cells were similar in both the malnourished and the control cattle. However, significantly lower numbers (P < 0.01) of circulating BoCD2+ cells, BoCD8+ cells, WC1+ gamma delta T cells and ACT2+ cells were found in the malnourished cattle. With the exception of inorganic phosphate, the changes in plasma biochemical parameters were unremarkable.

The role of nematode parasites in Soay sheep (Ovis aries L.) mortality during a population crash.

In early 1989, two-thirds of the Soay sheep population on St Kilda died over 12 weeks. Post-mortem examinations revealed emaciated carcasses and considerable nematode burdens, with protein-energy malnutrition as the probable cause of death. Haematological and blood biochemical changes in the sheep, as well as fecundity of gastrointestinal nematodes, suggested the hosts were immunosuppressed. In parallel, laboratory experiments in which Soay sheep on a high plane of nutrition were artificially infected with Ostertagia circumcincta, showed no clinical signs or mortality when supporting worm burdens similar to those recorded in dead sheep on St Kilda. Anthelmintic treatment of a group of animals increased daily survival rates in ewes and male lambs, although treated animals became re-infected as the 'crash' progressed. It is suggested that parasites contribute to mortality in malnourished hosts, exacerbating the effects of food shortage.

Repercussöes da desnutriçäo protéico-calórica sobre o colágeno, proteína total, RNA, DNA e peso pulmonar de ratas adultas / Effects of protein-caloric restriction on collagen, total protein, RNA, DNA and lung weight of adult female rat lungs

A restriçäo protéico-calórica em ratas adultas, durante 21 dias, acarretou queda significante nos pesos corporal e pulmonar desses animais, sem alterar significantemente o conteúdo de água e de DNA de seus pulmöes. A desnutriçäo protéico-calórica provocou, ainda, reduçäo significante no conteúdo pulmonar de RNA, proteína total e das relaçöes RNA/DNA e proteína total/DNA, indicando uma possível diminuiçäo da sintese protéica e do tamanho das células pulmonares respectivamente. As concentraçöes de RNA e proteína total foram semelhantes enquanto a de DNA foi signifivantemente superior nas ratas desnutridas. A desnutriçäo protéico-calórica näo afetou o conteúdo de hidroxiprolina do pulmäo enquanto ocasionou uma elevaçäo significante nas concentraçöes desse aminoácido, sugerindo a ocorrência de maior deposiçäo de colágeno ou a lentidäo de seu "turnover" em relaçäo aos das demais proteínas desse órgäo

Concentrations of serum constituents in cold-stressed calves from heifers fed inadequate protein and(or) energy.

A study with neonatal calves was conducted to determine the effects of maternal crude protein (CP) and(or) metabolizable energy (ME) malnutrition, cold stress (0 or 21 degrees C), and age on concentrations of selected serum constituents. For each of 2 yr, 60 artificially bred Angus heifers were assigned randomly to a 2 x 2 factorial nutritional plan 150 d before predicted parturition. The diets provided each heifer with either .32 or .96 kg/d of CP and 8.7 or 12.6 Mcal/d of ME. Blood samples were obtained from heifers at parturition and from their calves at birth and at 12, 24, 36, 48, and 72 h of age. Sera were analyzed for concentrations of blood urea nitrogen (BUN), creatinine (Creat), iron, total protein (TProt), alkaline phosphatase (AlkPhos), total bilirubin (TBil), and cholesterol (Chol). Mean correlations of these constituents in calf sera between 12-h adjacency intervals were high, but those between longer times (48 or 60 h) were low. Simple correlations of serum constituents between cows and calves at birth were low except for BUN (r = .578 and .295 for yr 1 and 2, respectively). There were significant main treatment effects for maternal CP consumption on BUN levels, for environmental temperature on BUN, Creat, and TBil levels, and for years on BUN, Creat, iron, and AlkPhos levels in calves. Significant polynomial relationships were found over hours of age for all variables. Blood urea N decreased in normal calves but remained relatively constant at a low level in deficient calves. Year x hour of age interactions occurred for iron, TProt, AlkPhos, TBil, and Chol. Protein x year x hour of age interactions were found for iron and Chol. These results suggest that random sampling times are not useful for decision making during the first 72 h after birth. Consideration must be given to multiple samples taken at specific calf ages, to environmental temperatures, and to maternal protein nutritional levels when interpreting calf blood sera data.

The use of parenteral nutrition in calves.

Parenteral nutrition has been an important adjunct to therapy of abdominal diseases in calves, with chronic diarrhea and wasting being the most common indication. Parenteral nutrition is administered on a short-term basis to prevent further protein-energy malnutrition in debilitated calves that cannot or will not consume adequate quantities of milk. Parenteral nutrition solutions consist of a protein source (amino acids) and energy sources (glucose and lipid emulsions), supplemented as needed with balanced electrolytes and vitamins. Complications due to PPN are rare, and it is the authors' clinical impression that survivability is enhanced when PPN is employed, although enhanced survivability was not demonstrated in one controlled experimental trial.

Protein-energy malnutrition in ruminants.

Protein-energy malnutrition occurs when feed is provided to ruminant livestock in insufficient quantity, quality, or both. The clinical syndrome that results from protein-energy malnutrition is not difficult to recognize, but it may be difficult to convince owners of the diagnosis. Development of clinical signs, such as recumbency and hypothermia, may occur rapidly owing to the sudden failure of homeostatic mechanisms that maintain the supply of cellular fuels. The ruminant is unique in its response to malnutrition because ruminal microorganisms become malnourished just as their host does. Ruminal maldigestion hastens the onset of clinical signs and makes recovery very difficult and prolonged. Clinical signs of PEM are similar in adult beef cattle, dairy cattle, sheep, and goats; however, the typical history of affected animals varies for each of these species. Neonatal ruminants may also be severely affected with PEM if they do not receive sufficient colostrum and milk. Definitive diagnosis of primary PEM requires necropsy of an affected animal. Diagnosis of PEM in an individual animal usually indicates a herd or flock problem that requires immediate attention. If the affected individual is already recumbent, then treatment will likely be difficult and unsuccessful. Changes in management of the herd or flock that involve ensuring adequate feed intake, minimizing cold and social stress, and meeting the animal's specific nutritional requirements will prevent PEM and maximize production.

Ontogeny of the immune response: effect of protein energy malnutrition in neonatal calves.

Immunocompetence of neonatal, Holstein bull calves fed for maximal growth (Control; n = 4) or protein energy malnutrition feeding (PEM; n = 4) for four weeks was assayed in vitro and in vivo. All calves exhibited elevated cortisol levels for ten days postnatally. At this time calves also were neutrophilic and lymphopenic. In addition lymphocyte function, as measured by lymphocyte proliferation and interleukin-2 activity, was reduced at this time as compared to older calves. After two weeks of protein energy malnutrition feeding, calves had significantly lower body weight, lymphocyte interleukin-2 activity and lymphocyte proliferation when compared with age-matched controls. Two weeks after protein energy malnutrition ration reversal, interleukin-2 activity and lymphocyte proliferation was comparable for both groups. There was no significant difference in serum cortisol concentration between control and protein energy malnutrition calves. The kinetics of the protein energy malnutrition group's primary humoral immune response was retarded, thus significantly lower antibody levels to K99 antigen were observed 8 to 12 days postimmunization. There was no significant difference between groups when comparing secondary response to K99 antigen.

Antibody responses in protein-energy restricted beef cows and their cold stressed progeny.

Antibody titers were measured in serum and colostral whey of pregnant beef cows immunized with tetanus toxoid and chicken red blood cells while being fed diets either restricted or nonrestricted in protein and/or metabolizable energy during the last 150 days of gestation. Serum antibody titers were also measured in the colostrum-fed, cold and noncold stressed progeny that were actively immunized with dinitrophenol conjugated to keyhole limpet hemocyanin. In general, there were no major or sustained differences in humoral immune responses to injection of tetanus toxoid or chicken red blood cells between cows fed diets that were adequate or restricted in protein or metabolizable energy. In the few cases where serum antibody titers to tetanus toxoid or chicken red blood cells differed (P less than 0.05) between adequately fed or restricted cows, the differences were no greater than twofold. Anti-chicken red blood cell titers were uniformly low (P less than 0.05) by a magnitude of two to threefold in colostral whey of cows restricted in protein and/or metabolizable energy when compared to titers in cows fed adequate amounts of protein and metabolizable energy. With one exception, neither maternal dietary restriction nor cold exposure had a major effect on the ability of the calves to absorb antitetanus toxoid and chicken red blood cell antibodies from colostrum. The humoral immune responses of all calves to injection of keyhole limpet hemocyanin and dinitrophenol were similar in magnitude.

In vitro incorporation of (U-C14)-glucose and (1-C14)-sodium acetate in peripheral nerves of malnourished young rhesus monkeys.

The effect of protein calorie malnutrition (PCM) on synthesis of lipids in peripheral nerves was studied by in vitro incorporation of (U-C14)-glucose and (1-C14)-sodium acetate. Ulnar and tibial nerves obtained from five young rhesus monkeys with PCM, five rehabilitated monkeys, and five control monkeys were incubated for 2 h with the radioactive precursors. Uptake of both radioactive precursors in whole peripheral nerves as well as myelin marker lipids was significantly decreased in animals with PCM. However, uptake returned to normal in rehabilitated monkeys.

[Effect of moderate protein deficiency on reproduction in the female rat and on the development of the pups until weaning]. / Effet d'une carence protéique modérée sur la reproduction chez la rate et sur le développement des jeunes jusqu'au sevrage.

Two groups of Wistar female rats were respectively fed ad libitum a standard stock diet containing 22 p. 100 protein (n = 93) and a diet containing 7.5 p. 100 protein (n = 189) for 8 weeks. They were mated with male rats of the same strain after 2 weeks of these diets. A small decrease (8 p. 100) in fecundity was observed but this moderate protein deprivation did not affect either the litter size (9.68 +/- 3.50 vs 9.61 +/- 3.69) or the percentage of stillborn pups (4.8 vs 4.9 p. 100). The postnatal mortality of the pups of deprived dams was much higher than that of pups from normal dams (11.2 vs 0.9 p. 100). During the suckling period, the 7.5 p. 100 protein diet did not cover the requirements of the dams. They lost 20 p. 100 of their weight, whereas the weight of the dams fed the 22 p. 100 protein diet remained stable. The weight deficit of the young rats born from deprived dams was about 10 p. 100 at birth but it rose to 50 p. 100 at weaning. During the gestation and suckling periods, the maternal body stores and tissues were mobilized to assure the growth of the young.

[Similarity of various clinical aspects of protein-calorie malnutrition to copper, zinc and cobalt deficiencies in ruminants]. / Similitude de certains aspects cliniques de la malnutrition protéino-énergétique avec les carences en cuivre, zinc et cobalt chez les ruminants.

Following clinical observations carried out on numerous herds in different countries the authors show that many similarities exist between proteino-energetic malnutrition and some symptoms of copper, zinc or cobalt deficiencies in ruminants. The probable origin of such a relationship is highlighted and the possible use of trace element therapy in livestock under malnutrition conditions is discussed.

Calorie-protein deficiencies and the immune response of the chicken I. Humoral immunity.

How dependent is the chicken on the calorie (C) and amino acid (AA) levels of the diet? This question was addressed by formulating diets that were two-thirds deficient in C and AA. Feeding the one-third C-one-third AA, one-third C and one-third AA diets for 5 weeks, in general reduced the primary response to 5% sheep red blood cells (SRBC) but not to .5% SRBC. The C/AA deficient birds were returned to a basal diet at 5 weeks and three weeks later reimmunized with SRBC. The secondary response, immunoglobulin G (IgG) titer, of the latter birds after 3 weeks on a basal diet was significantly lower than controls. Total serum protein was significantly depressed during the feeding of one-third AA diet. Transferrin, IgG, and IgM were not affected while albumin was significantly reduced. Gain in body weight was reduced in all groups receiving the C/AA deficient diets. Three weeks after repletion, all groups gained equally.