Some weeks later, as no clinical improvement was noted, the lemur was anesthetized to perform a more detailed examination. A big firm mass was revealed in the epigastrium by the abdominal palpation and it was confirmed by radiographs: a large radiopaque mass about 3 – 4 cm in diameter. Two days later a laparotomy was performed: the mass completely occluded the intestinal wall and was removed through intestinal resection and anastomosis . Consequently to this surgery, the subject showed a slight recovery from the illness then it became again anorexic and did not accept fragmented/homogenized food. Therefore the lemur was submitted to a second surgery , to which it did not survive. In this occasion internal organs were collected. Macroscopic evaluation of organs revealed: thickening and hypertrophy of the intestinal wall; presence of whitish sclerotic areas on the liver surface. The other organs did not show gross lesions.The intestine sample collected at first surgery revealed heavy changes of the intestinal wall in the area of the ring inspissation. The luminal surface was widely ulcerated, the tonaca propria was completely absent. There was a massive degeneration of the submucosal tonaca and a strong and exuberant connective new growth as well as a partial destruction of the muscular tonaca. The intestinal wall was heavily compromised and showed an intense lymphoid infiltrate, displaying a multifocal round and dense pattern. Sometimes these lymphocyte aggregates were perivascular. The cellular infiltrate involved a hyperplastic reactive stroma, composed of immature connective tissue, strongly vascularized.
The lymphoid cells were morphologically homogeneous. The nuclei were small , euchromatinic,round plastic pots with polygonal shape, and often presented indented membrane. They contained multiple, medium sized nucleoli and chromatin aggregates. The cytoplasm tent to be diaphanous and slightly reticular. The mitotic index was high. At the edge of the ring-shaped lesion there was a peculiar inflammatory feature.Plasma cells and lymphocytes infiltration in the lamina propria; impressive villous atrophy and crypt hyperplasia with decreased villous height-to-crypt depth ratio; pronounced increase in the number of intraepithelial lymphocytes and Figure 3. The primary lesion was identified as media cells intestinal lymphoma associated with enteritis, and the peripheral inflammatory pattern was similar to that described in the case of grade 4 Marsh-Oberhuber human coeliac disease . In this occasion the main internal organs were collected and processed for histology. The macroscopic evaluation of organs revealed: thickening and hypertrophy of the intestinal wall; presence of whitish sclerotic areas on the liver surface. The other organs did not show gross lesions. The histological evaluation of the intestine Haematoxylin-Eosin sections collected at two different levels at necroscopy, pointed out inflammatory alterations similar to those observed at the edge of the neoplastic lesion found in the first intestinal sampling. Intestinal specimens collected at two different levels of the intestine were similar. The villi were strongly atrophic and flattened and the mucosa assumed peculiar colonic-like features.
The epithelial cells showed a karyorectic multi-focal necrosis and intraepithelial lymphocytes were abundant. The tonaca propria displayed an intense lymphocyte/plasma cells infiltrate, as well as areas of necrosis. At the base of the muscularis mucosae a slight lymphocyte infiltration was detectable. The tunica muscularis and tunica serosa presented foci of lymphocyte infiltration. The histopathological data concerning the intestine could suggest the presence of a malabsorption related-enteritis similar to the human coelic disease . The intensity and the pattern of lymphocyte recruitment suggested the possible neoplastic progression of the lesion. The liver parenchyma presented an evident architecture destructuration of the lobules. Moreover, the lobules showed areas of multi-focal necrosis . Sometimes the necrotic areas were adjacent to the thickened Glisson membrane. An intense and diffuse infiltration of lymphoid cells was detectable, resembling those invading the tonaca propriaof the intestine. Associated to these features there were also evident areas of hemosiderin accumulation, considered as a common finding in zoo lemurs. Lung parenchyma presented a diffuse oedema associated to areas of destructive emphysema. The apical part of the lung revealed an area of acute pneumonia, with infiltration of neutrophilgranulocytes, involving the apical part of the lung. The spleen was morphologically intact but it was involved in the recruitment of a strong infiltrate composed by lymphoid cells similar to those observed in gut and liver.In order to identify the lineage of the neoplastic lymphocyte population and the inflammatory infiltrating lymphocytes, the histological sections were submitted to an immunoperoxidase based staining, using the following antibodies: polyc-lonal rabbit anti-human CD3 ; monoclonal mouse anti-human CD79α ; monoclonal mouse anti-human CD20 .
The CD3 antibody is specific for T cells and allows the immune detection of T lymphocytes derived neoplasms. The CD79α antibody is specific for B cells and allows the immune detection of neoplasms deriving from B lymphocytes at all stages of maturation. The CD20 antibody is specific for B cells precursors and mature B cells, excluding plasma cells. As positive controls, sections of human healthy lymph node, human skin characterized by lymphocyte infiltration and healthy lemur skin, were submitted to the same immuno histochemical procedure, using anti CD3, anti CD79α and anti CD20 as markers.In the intestine sections a relevant part of infiltrating lymphocytes was immuno reactive for the T-lymphocyte marker CD3 and Figure 3, revealing inflammatory features. Also numerous IELs were positive and a slight positivity was found also in the marginal part of the Auerbach plexus . The B-lymphocyte marker CD20 did not identify positive cells among the infiltrating lymphocytes. Only few granular cells localized in the epithelium were positive. The anti CD79 antibody was ineffective on lemur healthy spleen and due to this lack of cross-reactivity was not considered suitable to evaluate the immunophenotype of infiltrating cells. The intestine sections of the neoplastic lesion, when treated with the CD3 antibody, revealed the positivity of all the neoplastic lymphocytes, becoming more intense in the cells localized marginally to the main lesion. The CD20 antibody provided only a slight staining of some granular cells, as observed in the inflammatory lesions. The liver sections revealed a strong positivity of infiltrating cells to CD3, suggesting the presence of a T-lymphocyte population. IHC confirms the diagnosis of T-cells intestinal lymphoma and a peripheral coeliac-like enteritis.Few studies have been investigating lymphoma in primates and reports of this tumor in lemurs are extremely rare.
Findings from the current study are important as they provide useful information on lymphoma in the Lemur catta,highlighting an association between this tumor and coeliac disease. Basing on previous literature, humans with coeliac disease have been found to be at higher risk to develop small-bowel non-Hodgkin’s lymphoma. This study highlighted that similar association may exist also in lemurs, which are the most phylogenetically distant primates from our species. In particular, in human pathology enteropathy-type T-cell lymphoma is a recognized complication of gluten-sensitive enteropathy. Tumour cells probably derive from a subset of intraepithelial lymphocytes , as suggested by their ability to invade crypt epithelium. The routine application of immunohistochemical staining for lymphocyte markers such as CD3 has been proposed as a mean to better evaluate the number and distribution of IELs and the lymphocytes composing the lymphoma associated to the enteropathy. The immunophenotyping approach has been already proposed by Pye et al. in a lemur affected by intestinal lymphoma, resulting as a B cell lymphoma enriched with a T cell infiltrate. In our experience a relevant part of infiltrating lymphocytes in the intestine, most of the lymphocytes composing the intestinal neoplasm, and the liver lymphoid infiltrate, were immunoreactive for the T-lymphocyte marker CD3. These findings reinforce the diagnostic hypothesis proposed by the authors. In comparative terms, the role of dietary related disorders in captive lemurs should be considered as possible aetiology. Although the causative agent is still uncertain, a gluten-sensitive enteropathy should be considered. The possible sources of dietary gluten in captivity are biscuits, flours and other accidental foods. In 2005, although not allowed in the zoo, interactions between visitors and lemurs were frequent, due to the enclosure design. Although the lemur diet was made of fruits and vegetable and no gluten-food was provided, uncontrolled food items were given to the lemurs by the visitors. In 2010, all lemur groups were moved in a new area, in large naturalistic enclosure with no possibility for visitor-animal interactions.
Globally wheat is cultivated to meet the consumable food demand of the humanity . Being a primary source of nutrition for millions of individuals, wheat contributes 30% of the total grain production of the world. However, annual yield gain in crop productivity is prolonged to meet the future demands for plant-based products for the projected global population in 2050 . Globally, different kinds of stresses are the main threats for production of crops . Plants are immobile, for their growth and for survival they need nutrients and water from the soil. Previously it has been reported that around 20% of the world cultivated area is being pretentious by salinity . Soil salinity adversely affects crop production up to 60%, and its drastic effect is more pronounced in the perspective of climate change scenario . Deposition of more salts in the upper layer of soil poses highly stressful environment for plant development, which ultimately causes reduction in yield or plant death . In Pakistan, salinity affected the 6.3 106 ha of irrigated land . This makes the circumstance disturbing and it represents a danger of deficiency of food. Recently, much consideration is given to wheat improvement by joining both new and traditional crop breeding approaches along with advances in management practices to cope with salinity problem . Improvement through conventional breeding requires the exploitation of the existing genetic variation in the wheat crop for salt tolerance. Past examinations have uncovered large variation for salt resilience in wheat. Genetic diversity is also high in local and/or exotic material, hydroponic bucket which might be valuable for the improvement of salt-tolerance in future breeding plans. For this purpose, different methods were used which includes solution, sand, and pot cultures, saline raised beds, and field screening . Field evaluation may be sometime misleading or inaccurate as salinity intermingles with drought. In solution culture, supplements are promptly accessible to plant in differentiating with soil, where filtering or chelation of supplements to soil ruins their accessibility to plants. This strategy is reasonable due to the lower climatic variation , and it has been used by several researchers for genotype screening, especially under salinity .
Within the field evaluation, different elements create problems inside the assurance of salt tolerant genotypes which are lenient in one climate may not be lenient in another climate . Additionally, soil features like pH, sodicity and harmful components i.e., boron change from zone to region indeed from land to land . Changing field and climates, genotype climate associations should be analyzed cautiously for prevalent and trustworthy outcomes in salt affected regions. In wheat crop, root, shoot, and their biomass have moreover been detailed as principal characteristics which bestow salt resistance. Wheat researchers concluded that shoot biomass and plant biomass might be utilized for choice of salt-tolerant genotypes . Salt stress created a huge decrease in wheat plant biomass, and this decline was more clear at high levels of salt stress as compared to low levels . Since salt-tolerant genotypes had the option to hold their growth and had more biomass than the salt-susceptible ones. Increased biomass of resistant genotypes might be connected with their cap-potential to keep a preferable photosynthesis rate over the susceptible genotypes . In light of association and heritability, it transformed into presumed that lengths of root and shoot, their fresh and dry weights, have positive associations and high heritability, so these might be exceptionally gainful norms for settling on salt tolerant genotypes . Identification of hereditary variation is an essential of any breeding plan pointed toward creating salt-tolerant genotypes. This study was directed to portray the relationship of considered seedling parameters and to choose reasonable determination standards under both control and saline climates. The present study will provide novel breeding material to develop a salt tolerant genotype suitable for saline conditions.The current research was directed in the experimental area of University of Agriculture, Faisalabad , 31 260 N, 73 060 E, and 184.4 m above sea level .