1) The document discusses the process of cryopreservation of buffalo (Bubalus bubalis) semen, including collection, characteristics, diluents, cryoprotectants, freezing, thawing, and use for artificial insemination.
2) Key aspects of cryopreservation include using tris or lactose-based diluents, adding 6-7% glycerol as a cryoprotectant, slow cooling rates, and 2-6 hour equilibration periods.
3) Successful artificial insemination with frozen-thawed buffalo semen can achieve conception rates over 50%, but rates are typically lower than for cattle due to anatomical differences in female buffal
the liver is the central laboratory of a chicken’s body. It is
essential that this organ is kept in an excellent condition in
order to maintain a healthy bird. Understanding the metabolic
function and causes of disruptions in liver functions helps us
to provide the birds with the right feed and health treatment.
This is the first lecture in the series of lectures covering the complete syllabus of Veterinary Obstetrics for veterinary students, practitioners, and researchers. This lecture describes fertilization, maternal recognition of pregnancy, and implantation.
The science of synchronization of estrus and ovulation in females has made great strides.
Several protocols that allow producers to precisely schedule insemination of groups of females are available for fixed-time insemination in females.
the liver is the central laboratory of a chicken’s body. It is
essential that this organ is kept in an excellent condition in
order to maintain a healthy bird. Understanding the metabolic
function and causes of disruptions in liver functions helps us
to provide the birds with the right feed and health treatment.
This is the first lecture in the series of lectures covering the complete syllabus of Veterinary Obstetrics for veterinary students, practitioners, and researchers. This lecture describes fertilization, maternal recognition of pregnancy, and implantation.
The science of synchronization of estrus and ovulation in females has made great strides.
Several protocols that allow producers to precisely schedule insemination of groups of females are available for fixed-time insemination in females.
This lecture on veterinary obstetrics describes the placenta formation and its types in domestic animals. The lecture would be useful for students, practitioners, and researchers.
Protocol for Cryopreservation of snowtrout semenN K Agarwal
Schizothorax richardsonii is an indigenous snow-trout in the Himalayan region of Uttarakhand. It contributes a lot to the fish food basket in the hilly area of the state. In past few years snowtrout capture fishery has registered continuously decreasing trend due to over-exploitation and habitat destruction. For reestablishing its status and propagation of the species, first successful attempt was made to cryopreserve the semen of S. richardsonii to make all time availability of viable sperm to increase hatchery production for river ranching and culture practices.
To develop the cryopreservation protocol for S. richardsonii, initially semen quality was evaluated as per species trait and for sperm motility behaviour. Good quality semen samples (>75% motile sperm after activation) were pooled and frozen within 2 hours of collection. Four extenders and two cryoprotectants (CPA) were tested to cryopreserve the S. richardsonii semen. The extender-Mounib’s medium was found most successful. The DMSO proved better CPA than glycerol. Equilibration time was standardized as 45 min for DMSO and 60 min for glycerol. The semen is diluted with the different diluents (extender + cryoprotectant) before freezing. Two semen dilution ratios 1:4 and 1:10 were tested for better viability after cryopreservation. French medium straws of 0.5 ml capacity were used to store the semen in liquid nitrogen. Thawing temperature was standardized as 20-250C.
The cryopreservation success was evaluated by post-thaw motility and fertilizing ability of cryopreserved semen. Post-thaw motility percentage and duration was significantly reduced. Fertilizing capacity of 375 days old cryopreserved semen at 1:4 dilution ratio was almost equal to that of fresh semen (>98%). Semen cryopreserved in Mounib’s extender with 5% DMSO as CPA, even after 375 days had hatching rate-35.91 ±0.15% and fry survival rate 21.28 ±0.41% while those from fresh semen it was 52.97±0.26% and 30.43±0.50% respectively. Though the hatching and survival rates were significantly superior with fresh semen (P<0.001) than the cryopreserved semen, but these results are still encouraging. This developed cryopreservation protocol for S. richardsonii semen has ensured year round availability of viable sperm for optimal utilization in hatchery production. It would open up new perspective such as establishing genetic material reserves, sperm / gene bank for selective breeding, cross breeding and ex-situ conservation of fish germplasm.
This lecture on veterinary obstetrics describes the placenta formation and its types in domestic animals. The lecture would be useful for students, practitioners, and researchers.
Protocol for Cryopreservation of snowtrout semenN K Agarwal
Schizothorax richardsonii is an indigenous snow-trout in the Himalayan region of Uttarakhand. It contributes a lot to the fish food basket in the hilly area of the state. In past few years snowtrout capture fishery has registered continuously decreasing trend due to over-exploitation and habitat destruction. For reestablishing its status and propagation of the species, first successful attempt was made to cryopreserve the semen of S. richardsonii to make all time availability of viable sperm to increase hatchery production for river ranching and culture practices.
To develop the cryopreservation protocol for S. richardsonii, initially semen quality was evaluated as per species trait and for sperm motility behaviour. Good quality semen samples (>75% motile sperm after activation) were pooled and frozen within 2 hours of collection. Four extenders and two cryoprotectants (CPA) were tested to cryopreserve the S. richardsonii semen. The extender-Mounib’s medium was found most successful. The DMSO proved better CPA than glycerol. Equilibration time was standardized as 45 min for DMSO and 60 min for glycerol. The semen is diluted with the different diluents (extender + cryoprotectant) before freezing. Two semen dilution ratios 1:4 and 1:10 were tested for better viability after cryopreservation. French medium straws of 0.5 ml capacity were used to store the semen in liquid nitrogen. Thawing temperature was standardized as 20-250C.
The cryopreservation success was evaluated by post-thaw motility and fertilizing ability of cryopreserved semen. Post-thaw motility percentage and duration was significantly reduced. Fertilizing capacity of 375 days old cryopreserved semen at 1:4 dilution ratio was almost equal to that of fresh semen (>98%). Semen cryopreserved in Mounib’s extender with 5% DMSO as CPA, even after 375 days had hatching rate-35.91 ±0.15% and fry survival rate 21.28 ±0.41% while those from fresh semen it was 52.97±0.26% and 30.43±0.50% respectively. Though the hatching and survival rates were significantly superior with fresh semen (P<0.001) than the cryopreserved semen, but these results are still encouraging. This developed cryopreservation protocol for S. richardsonii semen has ensured year round availability of viable sperm for optimal utilization in hatchery production. It would open up new perspective such as establishing genetic material reserves, sperm / gene bank for selective breeding, cross breeding and ex-situ conservation of fish germplasm.
Human spermatozoa can tolerate a range of temperature. They are not very sensitive to damage caused by cooling possibly because of high membrane fluidity which is used as a technique to preserve spermatozoa in adverse conditions. cryopreservation technology has been a boon in every aspect of infertility & ART practice.
process of preserving an embryo at sub-zero temperatures, generally at an embryogenesis stage corresponding to pre-implantation.this presentation will clear you about slow freezing rapid freezing and vitrification method of embryo freezing.
Food processing, composition of milk, microorganism present, Different type of adulteration test (sugar, starch, Salt, urea), biochemical test (organoleptic, clot on boiling, alcohol, lactometer, fat determination, protein determination), different type of pasteurization, processing, spray drying, Standards of PFA, FSSAI, BIS (profile and regulations) & non food application of milk.
Successful management of delayed case of mastitis in cowsuren vet
In this we tried & eliminated pathogens with low antibiotics. The withdrawl period of milk is decreased. Treatment is of low cost. Easily applicable in field condition....
Cryopreservation and its application to aquaculture.pptxNarsingh Kashyap
What is Cryopreservation ?
Cryopreservation is a process where biological materials such as cells and tissues are preserved by cooling to very low temperatures, usually at -196°C (the temperature of liquid nitrogen), yet remain viable after later warming to temperatures above 0°C.
Cryopreservation in aquatic species goes back 65 years and began about the same time as similar research was performed in livestock (Blaxter 2011).
In India, NBFGR & CIFA are the primary organization carrying out fish sperm cryopreservation for long term gene banking (J. K. Jena 2012)
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
2. Storage of Buffalo
( Bubalus bubalis) Semen
Sansone, G., M. F. Nastri and A. Fabbrocini.
2000
3. "And indeed, for you in livestock is a lesson. We give
you (milk to) drink from that which is in their bellies,
and for you in them are numerous benefits, and from
them you eat."
(Surat Al-Mu'minūn : Verse 21)
Holy Quran says :
4. CRYOPRESERVATION
Minimize the Cross-contamination
by other cell lines
Benefits of Cryopreservation
The process of preserving the
sperm by freezing it in liquid
Nitrogen at -196˚c
Genotypic drift
Saving reagents, time
Increase the number of mobile
Normal spermatozoa
5. INTRODUCTION
• Buffalo population, 140 million
• Buffalo semen preserved like bovine semen
• Buffalo spermatozoa susceptible to hazards
• Diluents and Cryoprotectants
Milk Meat
IMPORTANCE
Leather
6. COLLECTION OF SEMEN
Semen is collected:
Trained Buffalo
Artificial vagina at 39˚C to 41˚C
Early Morning, before feeding
Take two ejaculates
Minimum interval (30 min)
11. Factors Influencing The Quality of
Semen
Photoperiod Age (3-5 years old)
Winter
Best quality of
semen
Spring
Best quality of
semen
Rainy Season
Better quality
Summer
Poor quality of
semen
Autumn
Poor quality of
Semen
Season
12. DILUENTS
• Decrease the
viscosity in
fluids
• Media in
which we
store semen
CRYOPROTECTANTS
• Reduced Microbial
Contamination
• Protect
Morphological
Changes
• Preserve Fertilizing
ability of sperm
EXTENDERS
• liquid diluent
• Preserve its
fertilizing
ability
• Provide
nutrients
• Antibiotics
13. Liquid Storage of Semen
Diluents
• 5˚c
• 24
Egg-Yolk Lactose
Tris
aminomethane
• 5˚c
• 72h
Glycerol
Add in TRIS for long
period storage
(72h)
5˚c
48h
5˚c
24h
Cow Milk
Egg-Yolk Citrate
5˚c
24h
Fig : Egg-Yolk Citrate
14. Frozen Storage of Semen
1- DILUENTS
Tris
• Highest post
thaw motility
• 54.1%
Lactose
• Better post
thaw motility
• 46.5%
Milk
Poor post
thaw motility
• 41.6%
15. 1) EGG YOLK:
Advantage
• Lecithin and Lipoprotein contents
• Preservation of lipoprotein sheath of sperm cell
• Protective against cold shock
2- CRYOPROTECTANTS
Egg yolk Glycerol
16.
17. Egg yolk in tris-glycerol-based freezing diluents
Beyond 5% did not show any significant improvement
Reducing the yolk level from 20%-5% did not
adversely affect the freezability of buffalo semen.
Concentration of Egg Yolk
18. Disadvantage Of Egg Yolk
Stimulates Enzyme
System of
spermatozoa
Deamination of
amino acids in egg
yolk
Yields Hydrogen
Peroxide
Toxic to spermatozoa
during storage under
aerobic conditions
19. 2) GLYCEROL
• Add in extenders
• Prevents the sperm from mortality
• Concentration is 6% to 7%
• 3%-2% or above 7% decreased the post thaw
motility of spermatozoa
20. High Molecular Weight Sugars :
Play Cryoprotective role
Alternating
permeability of
sperm membrane
Maintain
Electrolyte
balance
21. 3- Additives to freezing diluents
Certain Vitamins
Amino acids
Enzymes
As VITAMINS:
Best post thaw motility was obtained with Vitamin E
Satisfactory motility was obtained with Vitamin A
May improve the viability and
fertilizing ability of spermatozoa
22. 4- Antibiotics
• Penicillin (1000IU/ml) and Streptomycin
Commonly added to freezing diluents.
• But, Penicillin and neomycin are more effective.
• Microorganisms present in Buffalo Bull semen are
sensitive to these antibiotics.
Fig : Penicillin (1000) Fig: Neomycin
23. PROCESSING OF SEMENTransfer of Semen
Samples
Methods of Dilution
Cooling of Semen
Equilibration of Semen
Use of frozen thaw semen for
insemination
Thawing of Semen
Freezing of SemenFreezing of semen
24. 1- Transfer of semen samples
• Breeding farms far and delay in processing
• If semen processed in 1h after collection
No change in motility
Morphology of spermatozoa
• Sperms are placed in its own plasma
For 10-15 min
25. Agglutination
• Buffalo Semen may show agglutination
• Diluents are added to prevent semen from
agglutination.
Concentration of Diluents
• Motile spermatozoa are consistent for:
6h at 5˚c
0.3% Glycerol
26. 2- Methods of Dilution
Final Concentration
100-150 million sperms/ml
One-Step Method
Shorter Equilibration time (2-4hrs)
Two-Step Method
• Better result
• Long Equilibration time (6 hrs)
• 2nd diluent portion has higher concentration
27. Experiment by Fabbrocini et al., 1995
Glycerol
Commercial Milk Extenders to Semen
Two Variants are used as:
• 3% Glycerol
• 11% Glycerol1st
Variant
• 0.3% Glycerol
• 14.3% Glycerol2nd
Variant
2nd dil. 1hr before freezing
2nd dil. 1hr before freezing
28. 3- Cooling of semen
Cooling of semen
Slow cooling Rapid cooling
procedures procedures
Fig: liquid nitrogen storage container
29. In 1995, Dhami and Shani cooled semen:
• Initial Temperature is 30˚c
First Case: 30˚-5˚c in 120 min
Second Case: 30˚-5˚c in 60 min
• Better post thaw motility observed by slow
cooling
Slow cooling procedure is recommended
during pre- freezing processing of semen
30. 4- Equilibration of semen
Equilibration Duration
2-4hr.
While some scientist, recommended longer
equilibration time
6hr.
Water Buffalo Semen:
Stand at 5˚c
Not less than 2h
No longer than 6hr before freezing
31. 5- Freezing of semen
• After equilibration:
Semen is packed in mini straw (0.25 ml)
Frozen in liquid nitrogen tank.
Conception rate is
better in MINI Straws
than medium straws
32. Apparatus Used
• Straws are suspended in horizontal position.
• 1-4 cm above liquid nitrogen
• For 10-20min.
• After which, they are
immersed in liquid nitrogen
at -196˚c.
Simple Isotherm Box
34. 5- Freezing of semen (cont…)
Del Sorbo et al. examined two freezing procedures
i. Step-Wise freezing
ii. Continuous Freezing
Post thaw recovery rates better for
Step-Wise Freezing Curve
37. 6- Thawing of semen
Semen changes from a frozen solid
to a liquid by gradual warming
Fig: Semen in thermos with 35˚c water
to thaw
38. Characteristics of Thawed Spermatozoa
Forward motility
Membrane Integrity
Sperm-Oocyte interaction
Several parameters
Morphological state Physiological state
40. 7- Use of Frozen-Thawed Semen For
Insemination and Fertility Results
Poor conditions of hygiene
Detection of oestrus
Artificial insemination, also known as AI,
is a procedure which involves direct
insertion of semen into a female's womb.
Difficulties Regarding AI in Buffalo Cows
Teaser bulls detect oestrus
41. The low conception rate to AI in buffaloes due to:
Small size of uterine body
Semen could be introduced into uterine horn.
Low Conception Rate in Buffaloes
42. Pregnancy rate higher than 50% can be
regarded as a good result for
insemination with frozen-thawed
buffalo semen.
According to Vale:
