The aim of this study was to evaluate the effect of dried
L. leaves as a phytogenic mixture added to broiler feed on CD4
and CD8
T lymphocyte subpopulations, Newcastle disease virus (NDV) antibody ...titres, and the presence of
in faecal samples. The study was conducted on 100 male Ross 308 broilers, divided into four groups of 25 broilers, for a 42-day research period. The groups were housed separately in boxes on a litter of softwood shavings and were fed starter mixture from day 1 to day 21 and finisher mixture from day 22 to day 42. Industrial hemp (
) was grown in the Crkvina area, Croatia (latitude: 45°18'46.8″ N; longitude: 15°31'30″ E). The hemp leaves were manually separated, sun-dried, and ground to a powder. The mixture offered to the control group did not contain cannabis leaves, whereas the three experimental groups received mixtures containing mixed cannabis leaves in a quantity of 10 g/kg, 20 g/kg, or 30 g/kg (E_10, E_20, and E_30, respectively). The mean NDV antibody level was uniform in all study groups until post-vaccination day 14 and increased comparably with time. The percentage of CD4
and CD8
lymphocytes in the peripheral blood subpopulation showed statistically significant differences (
< 0.001) in the E_20 group as compared with the control group and both the E_10 and E_30 groups throughout the study period. As the broiler age increased, the CD4
-to-CD8
ratios also increased and were statistically significant (
< 0.0001) on day 42 in all experimental groups as compared to the control group. Comparing the control group with the experimental groups indicated that the bacterial count was lower in broiler groups having received feed with the addition of 20 g/kg and 30 g/kg
leaves. In conclusion, the
leaves were found to elicit a favourable immunomodulatory effect on cell-mediated and humoral immune responses in broilers via increased CD4
and CD8
lymphocyte subpopulations and higher CD4
:CD8
cell ratios, thus indicating enhanced immune function capacity. In addition,
leaves may have complementary effects on the broiler post-vaccination immune response, increase broilers' resistance to infectious diseases, reduce the effect of stress associated with vaccination, and improve broiler health and welfare.
Linseed is well known for abundant content of α-linolenic acid why it has been widely used as a source of n-3 fatty acid. Higher ratio of polyunsaturated fatty acids can increase lipid peroxidation. ...Selenium (Se) plays a key role in antioxidant enzymes and can therefore be regarded for use in attempts focused on protection of fatty acids from oxidation. The organic Se has been shown to have higher bioavailability compared to the inorganic selenium. The aim of this research was to determine the influence of dietary replacement of n-6 with n-3 polyunsaturated fatty acids on plasma and milk fatty acid composition in the dairy cows. The experiment was conducted on 30 dairy Holstein cows in the period from parturition until the 3rd week of lactation. Soybean meal from control (C) group was replaced with grounded linseed in the both experimental (LS and LS+Se) groups. In addition to that, in the 2nd experimental (LS+Se) group sodium selenite from premix was replaced with organic selenium. Blood and milk samples were collected at 21st day of lactation and fatty acids were determined using gas chromatography from it. Feeding dairy cows during transition period with linseed showed a positive impact on milk fatty acid composition by increasing (P<0.05) the proportion of n-3 fatty acids. Proportion of α-linolenic fatty acid increased (P<0.05) in the blood of both groups fed with linseed, but was significant (P<0.05) only in milk of LS+Se group. Such data indicated that the addition of organic selenium acted favourable on α-linolenic fatty acid increase in milk. During early postpartum period addition of linseed in meal of dairy cow’s increased (P<0.05) ratio of oleic acid, but in combination with organic source of selenium this increase was not evident (P>0.05).
Food safety, climate change, the emergence of infectious diseases, the ban on the use of antibiotics as growth promoters, and increasingly demanding intensive production are daily challenges for ...poultry production. A functional immune system is a prerequisite for animal health, and nutrition is one of the modulators of the immune system; therefore, the appropriate balance of nutrients is extremely important for the proper development and maintenance of the immune system of animals. The antimicrobial and immunomodulatory effects of phytobiotics are properties that make their use important as feed additives for poultry. Cannabis sativa L. contains many different compounds such as flavonoids, terpenes, and cannabinoids, each with different properties and effects. The effects of C. sativa seeds, essential oils, and cakes as feed additives for poultry have already been investigated, but the effect of C. sativa L. leaves as a feed additive on immunostimulatory and antibacterial activity has not. The results of this study show that C. sativa as a phytogenic additive to animal feed has a favourable antimicrobial and immunomodulatory effect in the production of broiler chickens. The aim of this study was to evaluate the effect of dried Cannabis sativa L. leaves as a phytogenic mixture added to broiler feed on CD4sup.+ and CD8sup.+ T lymphocyte subpopulations, Newcastle disease virus (NDV) antibody titres, and the presence of E. coli in faecal samples. The study was conducted on 100 male Ross 308 broilers, divided into four groups of 25 broilers, for a 42-day research period. The groups were housed separately in boxes on a litter of softwood shavings and were fed starter mixture from day 1 to day 21 and finisher mixture from day 22 to day 42. Industrial hemp (C. sativa) was grown in the Crkvina area, Croatia (latitude: 45°18′46.8″ N; longitude: 15°31′30″ E). The hemp leaves were manually separated, sun-dried, and ground to a powder. The mixture offered to the control group did not contain cannabis leaves, whereas the three experimental groups received mixtures containing mixed cannabis leaves in a quantity of 10 g/kg, 20 g/kg, or 30 g/kg (E_10, E_20, and E_30, respectively). The mean NDV antibody level was uniform in all study groups until post-vaccination day 14 and increased comparably with time. The percentage of CD4sup.+ and CD8sup.+ lymphocytes in the peripheral blood subpopulation showed statistically significant differences (p < 0.001) in the E_20 group as compared with the control group and both the E_10 and E_30 groups throughout the study period. As the broiler age increased, the CD4sup.+-to-CD8sup.+ ratios also increased and were statistically significant (p < 0.0001) on day 42 in all experimental groups as compared to the control group. Comparing the control group with the experimental groups indicated that the bacterial count was lower in broiler groups having received feed with the addition of 20 g/kg and 30 g/kg C. sativa leaves. In conclusion, the C. sativa leaves were found to elicit a favourable immunomodulatory effect on cell-mediated and humoral immune responses in broilers via increased CD4sup.+ and CD8sup.+ lymphocyte subpopulations and higher CD4sup.+:CD8sup.+ cell ratios, thus indicating enhanced immune function capacity. In addition, C. sativa leaves may have complementary effects on the broiler post-vaccination immune response, increase broilers’ resistance to infectious diseases, reduce the effect of stress associated with vaccination, and improve broiler health and welfare.
This study describes the introduction and spread of avian influenza A (H5N1) subtype in Croatia. Seventeen isolates were identified during the period from October 2005 to March 2006, all originating ...from wild birds. The full-length nucleotide sequence analysis of the hemagglutinin (HA) gene of seven representative isolates revealed that three distinct genetic strains involved in the outbreaks, implicating at least three independent introductions of the virus into Croatia during a relatively short period of time. All three genetic strains belonged to clade 2.2 (Qinghai-like viruses) and each strain displayed significant similarity to concurrent H5N1 viruses from other European countries. The dominant strain of the virus was present in all four affected areas and in all three bird species (mute swan, mallard, and black-headed gull), indicating cross-species transmission of the virus. Two other genetic strains were found, together with the dominant strain, only in a marsh at the Adriatic coast during late February and early March 2006, which could be associated with frozen water surfaces in the continental part of Croatia as well as in Eastern Europe in early 2006 and the movement of birds toward warmer areas. This is also the first isolation of highly pathogenic avian influenza virus of H5N1 subtype from apparently healthy black-headed gulls.
As immune responses to live and inactivated vaccines might differ, temporal responses of broiler chickens to vaccination were examined on the basis of the abundance in the circulating blood of gene ...transcripts of IFN-α, IFN-γ and IL-2, critical cytokines for immune responses. Blood samples were collected 6, 12 and 24 hours, and 7 and 14 days following vaccination with either live or inactivated Newcastle disease virus, La Sota strain, at 14 days of age, and the abundance of transcripts for each cytokine was assayed by real-time RT-PCR. Physiological saline and vaccine emulsion without viral antigen were administered to control groups for live and inactivated vaccine groups, respectively. The abundance of IFN-γ transcripts was elevated during the early times following vaccination and had reached baseline by the seventh day but the abundance of IFN-α transcripts remained elevated. Transcripts for neither IFN gene were detected in the control birds. The abundance of transcripts for each IFN was not different between the two vaccinated groups at any time. Transcripts for IL-2 were detected only in spleens from chicken embryos that had been inoculated with the live virus. The results show that cells stimulated to produce IFN-α and IFN-γ enter the circulating blood but those stimulated to produce IL-2 do not, or in very low number, and the IFN responses to both vaccines are the same.
In 2018, Croatia reported the largest outbreak of West Nile virus (WNV) infections as well as the re‐occurrence of human Usutu virus (USUV) infections. For the first time, fatal WNV and USUV ...infections were detected in wild birds. We analysed epidemiological characteristics and molecular epidemiology of WNV and USUV infections detected during 2018 transmission season. From April to November, 178 patients with neuroinvasive disease and 68 patients with febrile disease were tested for WNV and USUV. Viral RNA was detected in cerebrospinal fluid (CSF) and urine samples using a real‐time RT‐PCR. Positive samples were tested by nested RT‐PCR and nucleotide sequencing. IgM/IgG antibodies were detected in serum/CSF samples using ELISA with confirmation of cross‐reactive samples by virus neutralization test (VNT). WNV neuroinvasive disease was confirmed in 54 and WNV fever in seven patients from 10 continental Croatian counties. Areas affected in 2018 were those in which cases occurred in previous seasons, while in three areas human cases were reported for the first time. Phylogenetic analysis of six strains from patients residing in different geographic areas showed circulation of WNV lineage 2. In three patients, neuroinvasive USUV infection was confirmed by RT‐PCR or VNT. Sequence analysis of one detected strain revealed USUV Europe 2 lineage. During the same period, a total of 2,574 horse and 1,069 poultry serum samples were tested for WNV antibodies using ELISA. Acute asymptomatic WNV infection (IgM antibodies) was documented in 20/0.7% horses. WNV IgG antibodies were found in 307/11.9% horses and in 125/12.7% poultry. WNV RNA was detected in two goshawks and USUV RNA was detected in one blackbird from north‐western Croatia. In the Zagreb area, 3,670 female mosquitoes were collected. One Culex pipiens pool collected in July tested positive for USUV RNA. Our results highlight the importance of continuous multidisciplinary ‘One health’ surveillance of these emerging arboviruses.
A total of 142 black-headed gulls (BHGs), Chroicocephalus ridibundus, were captured during February and March 2009 at the Zagreb city rubbish dump (45.45 N 16.01 E) in order to collect cloacal swabs ...and serum samples. Cloacal swabs were tested by virus isolation in embryonated chicken eggs, which resulted in isolation of one avian influenza virus (AIV) that was of the H16 subtype. The collected sera were tested by blocking ELISA for avian influenza antibodies, resulting in 28.2% positive samples, which were retested by haemagglutination inhibition (HI) using H5 and H7 subtype antigens. Only one serum sample was positive for H5 and none for H7 antibodies. Statistically, no significant difference was found between the ages of AIV seropositive birds (chi square = 2.08, df = 1, P = 0.15). In contrast, regarding seroprevalence in different months of capture, a higher proportion of positive gulls was found during March than during February (chi square = 4.53, df = 1, P = 0.03), especially in younger birds (chi square = 7.67, df = 1, P = 0.006). This finding suggests that BHGs might contract AIIV infection during their aggregations in large numbers on rubbish dumps or similar feeding sites during the winter. Although only one off the 142 tested birds was positive for H5 subtype antibodies, this finding cannot be neglected seeing that apparently healthy BHGs can carry highly pathogenic AIV of the H5N1 subtype. Nevertheless, the results off our study have shown that BHGs are more often infected with other AIIV subtypes and therefore are most likely not the primary carriers of H5 AIV. To our knowledge this is the first detection off H16 AIIV in Croatia. Key words: avian influenza, H5 subtype, gulls, Chroicocephalus ridnibundus Tijekom veljace i ozujka 2009. na gradskom odlagalistu otpada u Zagrebu (45.45 N 16.01 E) uhvacena su 142 rijecna galeba ((Chroicocephalus ridibundus')) u cilju prikupljanja brisova necisnica i uzoraka krvi. Brisovi necisnica su pretrazeni izdvajanjem virusa u embrioniranim kokosjim jajima sto je rezultiralo izdvajanjem virusa influence ptica koji je bio podtipa H16. Prikupljeni uzorci seruma su pretrazeni imunoenzimnim testom za protutijela virusa influence ptica od kojih je 28,2% bilo pozitivno. Pozitivni uzorci su dodatno pretrazeni inhibicijom hemaglutinacije s antigenima podtipa H5 i H7 medu kojima je samo jedan bio pozitivan na H5 i nijedan na H7 protutijela. Statisticki nije ustanovljena znacajna razlika medu skupinama razlicite dobi seropozitivnih ptica (chi square = 2,08, df = 1, P = 0,15). Nasuprot tome, seroprevalencija u razlicitim mjesecima lova je imala veci udio tijekom ozujka nego tijekom veljace (((2 = 4,53, df = 1, P = 0,03), narocito u mladih ptica (3(2 = 7,67, df = 1, P = 0,006). Ovakav nalaz ukazuje na mogucu infekciju rijecnih galebova tijekom njihovoga brojnog okupljanja na odlagalistima otpada ili slicnim mjestima za hranjenje tijekom zime. Iako je samo jedna od 142 ptice bila pozitivna na protutijela za H5 podtip, nalaz se ne smije zanemariti znajuci da naizgled zdravi rijecni galebovi mogu nositi virus jako patogene influence podtipa H5N1. Unatoc tome, rezultati naseg istrazivanja pokazuju da rijecni galebovi cesce bivaju inficirani drugim podtipovima virusa influence ptiica te stoga najvjerojatnije nisu primarni nosioci virusa influence podtipa H5. Sukladno nasim spoznajama, ovo je prvi dokaz virusa influencce podtipa H16 u Hrvatskoj. Kljucne rijeci: influenca ptica podtip H5, galebovi, Chroicocephalus ridibundus
Zagorski puran kao hrvatska autohtona pasmina sa zaštitom zemljopisnog podrijetla i zaštićenom oznakom izvornosti sve se više uzgaja na obiteljskim poljoprivrednim gospodarstvima. Tradicionalni način ...držanja zagorskog purana na otvorenom, uz često nedovoljno prostora u objektima kao i izmetom, hranom i otpadom često zaprljani manji ispusti, način hranjenja uz neodržavanje higijene hranilica i pojilica i uzgoj raznovrsne peradi na istom prostoru, u prvom redu kokoši, predispozicija su za pojavu bolesti poput mikoplazmoze i histomonijaze. U takvim uvjetima česta je i sekundarna bakterijska infekcija bakterijom Escherichia coli. U radu smo dali osnovne smjernice uzgoja zagorskog purana, kao i pregled najznačajnijih bolesti kojima pogoduje tradicionalni način uzgoja zagorskog purana ustvrđenih tijekom dvogodišnjeg istraživanja na obiteljskim poljoprivrednim gospodarstvima Krapinsko- zagorske županije. Praćeno je ukupno 46 uzgoja s preko 10000 uzgojenih purana godišnje. U 22 % obiteljskih poljoprivrednih gospodarstava ustvrđen je zarazni sinusitis, a u 11 % uzgoja histomonijaza. Sekundarna bakterijska infekcija bakterijom E. coli dokazana je u 10 uzgoja zahvaćenih mikoplazmom ili histomonijazom. Povećanom uginuću i štetama u gospodarstvima vjerojatno je pridonijela i učestala, nasumična, nekontrolirana i nepotrebna uporaba antimikrobnih lijekova koji stvaraju rezistenciju bakterija. Uz zaštitu zagorskog purana kao hrvatskog izvornog proizvoda, od velike je važnosti provoditi i sustavni veterinarski nadzor kako bi se prevenirao razvoj bolesti te unaprijedio i povećao uzgoj ove važne hrvatske peradi.
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